Supplemental File 1: Laboratory procedures of tissue preparation, microarray and qPCR
analyses
Tissue preparation and microarray analysis
Tissue specimens at the margin of resection were obtained during surgery. All specimens were
immediately rinsed with cold diethyl pyrocarbonate-treated phosphate-buffered saline, snapfrozen in liquid nitrogen and stored at -80ºC. RNA was extracted using TRIZOL reagent (Life
Technologies, Gaithersburg, MD) and RNeasy mini kit (QIAGEN, GmbH, Hilden). Total RNA
(100 ng) was amplified and labeled using the Ambion whole-transcript expression kit (Life
Technologies) and Affymetrix WT sense target labeling kit (Affymetrix Inc., Santa Clara, CA).
Affymetrix GeneChip Human Gene 1.0 ST arrays were hybridized with 540 ng of labeled sense
DNA, washed, and stained using the FS450 Fluidic Station (Affymetrix). Fluorescent signal
intensities of each stained chip were captured by the GeneChip Scanner 3000 7G System
(Affymetrix).
Expression arrays were analyzed using the Partek Genomics Suite v6.5 software (Partek,
St. Louis, MO). Probeset signals were normalized with the Robust Multi-array Average (RMA)
method and log2-transformed. The three-group (NEC versus Surg-CTL, SIP versus Surg-CTL,
and NEC versus SIP) analysis was performed and the gene list was generated using the criteria of
(i) >2-fold differences and (ii) P<0.05 between NEC or SIP and Surg-CTL. The microarray data
sets were submitted to Gene Expression Omnibus database (GSE46619).
1
Gene ontology and network analysis
The web-based tool (Database for Annotation, Visualization, and Integrated Discovery, DAVID)
was used to identify enriched Gene Ontology (GO) biological processes among the differentially
expressed genes (NEC versus Surg-CTL and SIP versus Surg-CTL). In addition, subgroup
analysis was performed on categories of “epithelial cell proliferation” and “smooth muscle
proliferation”, as indicators of tissue viability. The MetaCore Analysis Suite (GeneGo-Thomson
Reuters, St. Joseph, MI) was used to map differentially expressed networks between the NEC
and SIP datasets. The networks listed were annotated with specific biologic functions based on
GO processes and ranked according to differences between NEC and SIP. For network analysis
of validated targets, we used multiple algorithms to map direct gene-gene interaction, traced
pathways and the shortest path for interactions.
Validation of target mRNA expression levels by qPCR
Gene expression levels of selected targets in NEC, SIP, Surg-CTL and Fetal-CTL tissues were
quantified by qPCR using pre-designed TaqMan assays (Life Technologies). A subgroup of
preterm infants [NEC (n=12), SIP (n=11) and Surg-CTL (n=9)] with small intestinal specimens
were further analyzed separately.
Total RNA (500 ng) was used to synthesize cDNA with the high capacity cDNA
synthesis kit (Life Technologies). The qPCR reaction was performed in duplicate with cDNA
(0.2 µL), 2 x TaqMan Gene Expression Master Mix (10 µL), and specific primers and probes
(final volume 20 µL). Amplification was performed for 45 cycles with denaturation at 95°C for
15 sec, and annealing extension at 60°C for 1 min. The emission intensity was detected by the
2
ABI 7300 Real-Time PCR System (Life Technologies). The average threshold cycles (Ct) were
used to calculate the expression ratios relative to glyceraldehyde 3-phosphate dehydrogenase
(GAPDH; Life Technologies).
Supplemental Figure 1. Legend for object symbols of the Metacore network
The legend for the object symbols shown in Figures 2, 3, 4 and Supplemental Figure 2 are listed.
These objects include enzymes, channels/transporters, receptors, generic classes, and G protein
adaptors and regulators.
Supplemental Figure 2. Interacting network of angiogenesis and extracellular matrix
(ECM) remodeling-related genes in NEC
A biological network of dysregulated target genes in the functional categories of angiogenesis
and ECM remodeling in NEC tissues was constructed based on the MetaCore software using the
direct interaction algorithm. Expression changes are demonstrated as red and blue color circles,
representing up- and down-regulation. The gene symbol or name of each target gene is shown
next to the object. Green, red and grey arrows between target genes represent positive, negative
and unspecified interactions, respectively. The effects of interaction between genes are indicated
as binding (B), cleavage (C) and transcriptional regulation (TR).
3
Supplemental Figure 1. Legend for object symbols of the Metacore network
4
Supplemental Figure 2. Interacting network of angiogenesis and extracellular matrix
(ECM) remodeling-related genes in NEC
5
Supplemental Table 1. Quantitative PCR assays and reference sequence numbers of the
selected genes
Gene
Angiogensis
ACE
ACE2
ANGPTL4
Assay ID
Reference Sequences
Hs00174179_m1
Hs01085333_m1
Hs01101127_m1
FLT1
Hs01052961_m1
GJA5
HBEGF
Hs00270952_s1
Hs00181813_m1
NR1H4
Hs01026590_m1
THBS1
Hs00962908_m1
NM_001178057.1;NM_000789.3;NM_152830.2
NM_021804.2
NM_001039667.1;NM_139314.1
NM_001160031.1;NM_001160030.1;NM_002019.4;
NM_001159920.1
NM_181703.2;NM_005266.5
NM_001945.2
NM_001206977.1;NM_001206978.1;NM_00120699
3.1;NM_001206979.1;NM_005123.3;NM_00120699
2.1
NM_003246.2
Arginine Metabolism
ARG1
Hs00968978_m1
CPS1
Hs00157048_m1
OAT
Hs00236852_m1
OTC
Hs00166892_m1
NM_000045.3;NM_001244438.1
NM_001122633.2;NM_001122634.2; NM_001875.4
NM_000274.3;NM_001171814.1
NM_000531.5
Cell Adhesion and Chemotaxis
CCL20
Hs01011368_m1
CD44
Hs01075861_m1
CXCL5
CXCR1
ICAM1
Hs01099660_g1
Hs01921207_s1
Hs00164932_m1
NM_004591.2;NM_001130046.1
NM_001001392.1;NM_001202555.1;NM_00100138
9.1;NM_001202556.1;NM_000610.3;NM_00100139
1.1;NM_001001390.1
NM_002994.3
NM_000634.2
NM_000201.2
ECM Remodeling
MMP1
Hs00899658_m1
MMP9
Hs00957562_m1
PLAUR
Hs00958880_m1
SERPINB2
Hs01010736_m1
NM_002421.3;NM_001145938.1
NM_004994.2
NM_001005377.2;NM_002659.3;NM_001005376.2
NM_001143818.1;NM_002575.2
SERPINE1
TFPI2
TIMP1
BC010860.1 (GenBank)
NM_006528.2
NM_003254.2
Hs01126606_m1
Hs00197918_m1
Hs00171558_m1
Hypoxia and Oxidative Stress
6
HIF1A
HMOX1
Hs00153153_m1
Hs01110250_m1
PRKCZ
Hs00177051_m1
PTGS2
SOD2
Hs00153133_m1
Hs00167309_m1
NM_181054.2;NM_001530.3;NM_001243084.1
NM_002133.2
NM_001242874.1;NM_001033581.1;NM_00103358
2.1;NM_002744.4
NM_000963.2
NM_000636.2;NM_001024465.1;NM_001024466.1
Inflammation
CD14
Hs02621496_s1
FOSL1
IL10
IL1B
IL6
IL8
MEFV
NFKB2
NLRC4
Hs04187685_m1
Hs00961622_m1
Hs01555410_m1
Hs00985639_m1
Hs00174103_m1
Hs00925524_m1
Hs01028901_g1
Hs00892666_m1
NLRP3
Hs00918082_m1
NOD2
TLR2
TLR4
TNF
TREM1
Hs00223394_m1
Hs00152932_m1
Hs01060206_m1
Hs01113624_g1
Hs00218624_m1
TYROBP
Hs00182426_m1
Muscle Contraction
Hs00154286_m1
CASQ2
GALR2
Hs00605839_m1
GNA11
Hs01588833_m1
KCNMA1
Hs01119498_m1
MYH11
Hs00224610_m1
MYLK
Hs00364926_m1
MYOM1
Hs00187003_m1
NM_000591.3;NM_001040021.2;NM_001174104.1;
NM_001174105.1
NM_005438.3
NM_000572.2
NM_000576.2
NM_000600.3
NM_000584.3
NM_000243.2;NM_001198536.1
BC002844.2 (GenBank)
NM_001199139.1;NM_021209.4;NM_001199138.1
NM_001127461.2;NM_001243133.1;NM_183395.2;
NM_001127462.2;NM_001079821.2;NM_004895.4
NM_022162.1
NM_003264.3
NM_138554.4
NM_000594.2
NM_018643.3
NM_198125.2;NM_003332.3;NM_001173514.1;NR
_033390.1;NM_001173515.1
NM_001232.3
NM_003857.2
NM_002067.2
NM_001014797.2;NM_001161352.1;NM_002247.3;
NM_001161353.1
NM_002474.2;NM_001040113.1;NM_001040114.1;
NM_022844.2
NM_053025.3;NM_053026.3;NM_053031.2;NM_05
3027.3;NM_053028.3;NM_053032.2
NM_003803.3;NM_019856.1
7
Supplemental Table 2. Microarray data analysis on biological processes of differentially
expressed transcripts between NEC or SIP and Surg-CTL tissues
NEC vs. Surg-CTL (upregulated)
Response to wounding
Defense response
Inflammatory response
Immune response
Chemotaxis
Taxis
Locomotory behavior
Behavior
Response to organic substance
Response to bacterium
Count
110
112
79
110
46
46
50
60
75
35
%
15.11
15.38
10.85
15.11
6.32
6.32
6.87
8.24
10.30
4.81
P-value
1.06E-46
9.12E-42
3.08E-38
2.02E-35
1.90E-25
1.90E-25
1.98E-18
1.86E-14
3.42E-13
6.91E-13
NEC vs. Surg-CTL (downregulated)
Digestion
Steroid metabolic process
Fatty acid metabolic process
Oxidation reduction
Lipid catabolic process
Lipid localization
Carbohydrate transport
Lipid biosynthetic process
Lipid transport
Intestinal absorption
Count
30
44
37
78
31
29
17
45
27
9
%
2.52
3.70
3.11
6.57
2.61
2.44
1.43
3.79
2.27
0.76
P-value
6.44E-14
5.04E-13
3.97E-09
9.21E-09
2.32E-07
3.22E-07
5.64E-07
5.83E-07
7.58E-07
1.68E-06
SIP vs. Surg-CTL (upregulated)
Defense response
Response to wounding
Immune response
Response to bacterium
Inflammatory response
Defense response to bacterium
Chemotaxis
Taxis
Regulation of angiogenesis
Positive regulation of cell division
Count
33
24
24
14
16
10
11
11
7
6
%
25.58
18.60
18.60
10.85
12.40
7.75
8.53
8.53
5.43
4.65
P-value
3.43E-18
1.81E-11
3.24E-09
3.80E-09
3.80E-08
2.41E-07
5.31E-07
5.31E-07
1.00E-05
1.31E-05
SIP vs. Surg-CTL (downregulated)
Excretion
Muscle contraction
Muscle system process
Digestion
Count
6
8
8
6
%
3.11
4.15
4.15
3.11
P-value
2.40E-04
0.001
0.001
0.002
8
Immune response
16
8.29
0.003
Digestive system process
4
2.07
0.004
Negative regulation of multicellular organismal process 7
3.63
0.005
Metal ion transport
12
6.22
0.005
Regulation of smooth muscle contraction
4
2.07
0.006
Cation transport
13
6.74
0.007
The top 10 significant GO biological process terms are listed. NEC n=5, SIP n=5 and
Surg-CTL n=4.
9
Supplemental Table 3. Microarray data analysis on the most differentially affected
networks between NEC or SIP and Surg-CTL tissues
P-value
NEC
SIP
1.23E-25
4.29E-13
1.52E-08
9.27E-06
4.09E-03
2.12E-03
4.32E-08
1.18E-02
6.56E-03
NS
NS
NS
Inflammation and Immune Response
Neutrophil activation
Innate inflammatory response
Histamine signaling
TREM1 signaling
Kallikrein-kinin system
Complement system
Amphoterin signaling
Protein C signaling
IgE signaling
Jak-STAT Pathway
IL-4 signaling
IL-10 anti-inflammatory response
Interferon signaling
NK cell cytotoxicity
Inflammasome
MIF signaling
Phagocytosis
Phagosome in antigen presentation
Antigen presentation
Th17-derived cytokines
1.07E-09
1.01E-08
1.94E-06
3.14E-06
8.55E-05
6.65E-05
9.86E-09
2.52E-04
4.48E-04
2.62E-04
1.20E-04
4.36E-05
1.36E-04
2.60E-03
1.89E-02
3.25E-03
3.77E-08
9.39E-05
3.69E-05
1.26E-07
NS
1.53E-02
NS
NS
NS
NS
3.56E-05
NS
NS
NS
2.50E-02
5.96E-03
6.18E-03
NS
NS
NS
NS
NS
NS
1.57E-04
Muscle contraction
NS
1.02E-04
Development
Regulation of angiogenesis
Regulation of epithelial-to-mesenchymal transition
1.67E-06
2.43E-04
1.00E-02
NS
Proteolysis
ECM remodeling
Connective tissue degradation
8.69E-05
4.44E-08
1.51E-07
1.37E-06
Process Networks
Chemotaxis and Cell Adhesion
Chemotaxis
Platelet-endothelium-leucocyte interactions
Cell-matrix interactions
Leucocyte chemotaxis
Cell junctions
Glycoconjugates
10
Apoptosis
Anti-Apoptosis mediated by external signals via 2.55E-04
PI3K/AKT
Anti-Apoptosis mediated by external signals via MAPK 1.73E-02
and JAK/STAT
Anti-apoptosis mediated by external signals via NF-kB 1.85E-02
1.48E-02
Response to Hypoxia and Oxidative Stress
NS, not significant
11
3.18E-02
NS
NS
NS
Supplemental Table 4. Summary of target gene names and functions
Gene
Gene name
ACE
angiotensin I
converting enzyme
(peptidyl-dipeptidase
A) 1
ACE2
angiotensin I
converting enzyme
(peptidyl-dipeptidase
A) 2
ANGPTL4
angiopoietin-like 4
FLT1
fms-related tyrosine
kinase 1
GJA5
gap junction protein,
alpha 5, 40kDa
HBEGF
heparin-binding EGFlike growth factor
Summary
This gene encodes an enzyme involved in catalyzing the conversion of angiotensin I into a
physiologically active peptide angiotensin II. Angiotensin II is a potent vasopressor and
aldosterone-stimulating peptide that controls blood pressure and fluid-electrolyte balance.
This enzyme plays a key role in the renin-angiotensin system. (provided by RefSeq, May
2010)
SwissProt: Q9BYF1 # Carboxypeptidase which converts angiotensin I to angiotensin 1-9, a
peptide of unknown function, and angiotensin II to angiotensin 1-7, a vasodilator. Also able to
hydrolyze apelin-13 and dynorphin-13 with high efficiency. May be an important regulator of
heart function. In case of human coronaviruses SARS and HCoV-NL63 infections, serve as
functional receptor for the spike glycoprotein of both coronaviruses.
This gene is a member of the angiopoietin/angiopoietin-like gene family and encodes a
glycosylated, secreted protein with a fibrinogen C-terminal domain. This gene is induced
under hypoxic conditions in endothelial cells and is the target of peroxisome proliferation
activators. The encoded protein is a serum hormone directly involved in regulating glucose
homeostasis, lipid metabolism, and insulin sensitivity and also acts as an apoptosis survival
factor for vascular endothelial cells. (provided by RefSeq, Jul 2008)
This gene encodes a member of the vascular endothelial growth factor receptor (VEGFR)
family. VEGFR family members are receptor tyrosine kinases (RTKs) which contain an
extracellular ligand-binding region with seven immunoglobulin (Ig)-like domains, a
transmembrane segment, and a tyrosine kinase (TK) domain within the cytoplasmic domain.
This protein binds to VEGFR-A, VEGFR-B and placental growth factor and plays an
important role in angiogenesis and vasculogenesis. (provided by RefSeq, May 2009)
This gene is a member of the connexin gene family. The encoded protein is a component of
gap junctions, which are composed of arrays of intercellular channels that provide a route for
the diffusion of low molecular weight materials from cell to cell. (provided by RefSeq, Jul
2008)
SwissProt: Q99075 # Growth factor that mediates its effects via EGFR, ERBB2 and ERBB4.
Required for normal cardiac valve formation and normal heart function. Promotes smooth
muscle cell proliferation. May be involved in macrophage-mediated cellular proliferation. It is
mitogenic for fibroblasts, but not endothelial cells. It is able to bind EGF receptor/EGFR with
12
NR1H4
nuclear receptor
subfamily 1, group H,
member 4
THBS1
thrombospondin 1
ARG1
arginase, liver
CPS1
carbamoyl-phosphate
synthase 1,
mitochondrial
OAT
ornithine
aminotransferase
higher affinity than EGF itself and is a far more potent mitogen for smooth muscle cells than
EGF. Also acts as a diphtheria toxin receptor.
SwissProt: Q96RI1 # Ligand-activated transcription factor. Receptor for bile acids such as
chenodeoxycholic acid, lithocholic acid and deoxycholic acid. Represses the transcription of
the cholesterol 7-alpha-hydroxylase gene (CYP7A1) through the induction of NR0B2 or
FGF19 expression, via two distinct mechanisms. Activates the intestinal bile acid-binding
protein (IBABP). Activates the transcription of bile salt export pump ABCB11 by directly
recruiting histone methyltransferase CARM1 to this locus.
The protein encoded by this gene is a subunit of a disulfide-linked homotrimeric protein. This
protein is an adhesive glycoprotein that mediates cell-to-cell and cell-to-matrix interactions.
This protein can bind to fibrinogen, fibronectin, laminin, type V collagen and integrins alphaV/beta-1. This protein has been shown to play roles in platelet aggregation, angiogenesis, and
tumorigenesis. (provided by RefSeq, Jul 2008)
Arginase catalyzes the hydrolysis of arginine to ornithine and urea. At least two isoforms of
mammalian arginase exist (types I and II) which differ in their tissue distribution, subcellular
localization, immunologic crossreactivity and physiologic function. The type I isoform
encoded by this gene, is a cytosolic enzyme and expressed predominantly in the liver as a
component of the urea cycle. Inherited deficiency of this enzyme results in argininemia, an
autosomal recessive disorder characterized by hyperammonemia. Two transcript variants
encoding different isoforms have been found for this gene. (provided by RefSeq, Sep 2011)
The mitochondrial enzyme encoded by this gene catalyzes synthesis of carbamoyl phosphate
from ammonia and bicarbonate. This reaction is the first committed step of the urea cycle,
which is important in the removal of excess urea from cells. The encoded protein may also
represent a core mitochondrial nucleoid protein. Three transcript variants encoding different
isoforms have been found for this gene. The shortest isoform may not be localized to the
mitochondrion. Mutations in this gene have been associated with carbamoyl phosphate
synthetase deficiency, susceptibility to persistent pulmonary hypertension, and susceptibility
to venoocclusive disease after bone marrow transplantation. (provided by RefSeq, May 2010)
This gene encodes the mitochondrial enzyme ornithine aminotransferase, which is a key
enzyme in the pathway that converts arginine and ornithine into the major excitatory and
inhibitory neurotransmitters glutamate and GABA. Mutations that result in a deficiency of
this enzyme cause the autosomal recessive eye disease Gyrate Atrophy. Alternatively spliced
transcript variants encoding different isoforms have been described. Related pseudogenes
13
OTC
ornithine
carbamoyltransferase
CCL20
chemokine (C-C motif)
ligand 20
CD44
CD44 molecule (Indian
blood group)
CXCL5
chemokine (C-X-C
motif) ligand 5
CXCR1
chemokine (C-X-C
motif) receptor 1
ICAM1
intercellular adhesion
molecule 1
MMP1
matrix
metallopeptidase 1
(interstitial
collagenase)
have been defined on the X chromosome. (provided by RefSeq, Jan 2010)
This nuclear gene encodes a mitochondrial matrix enzyme. Missense, nonsense, and
frameshift mutations in this enzyme lead to ornithine transcarbamylase deficiency, which
causes hyperammonemia. Since the gene for this enzyme maps close to that for Duchenne
muscular dystrophy, it may play a role in that disease also. (provided by RefSeq, Jul 2008)
SwissProt: P78556 # Chemotactic factor that attracts lymphocytes and, slightly, neutrophils,
but not monocytes. Inhibits proliferation of myeloid progenitors in colony formation assays.
May be involved in formation and function of the mucosal lymphoid tissues by attracting
lymphocytes and dendritic cells towards epithelial cells. C-terminal processed forms have
been shown to be equally chemotactically active for leukocytes. Possesses antibacterial
activity E.coli ATCC 25922 and S.aureus ATCC 29213.
SwissProt: P16070 # Receptor for hyaluronic acid (HA). Mediates cell-cell and cell-matrix
interactions through its affinity for HA, and possibly also through its affinity for other ligands
such as osteopontin, collagens, and matrix metalloproteinases (MMPs). Adhesion with HA
plays an important role in cell migration, tumor growth and progression. Also involved in
lymphocyte activation, recirculation and homing, and in hematopoiesis. Altered expression or
dysfunction causes numerous pathogenic phenotypes. Great protein heterogeneity due to
numerous alternative splicing and post-translational modification events.
SwissProt: P42830 # Involved in neutrophil activation. In vitro, ENA-78(8-78) and ENA78(9-78) show a threefold higher chemotactic activity for neutrophil granulocytes.
SwissProt: P25024 # Receptor to interleukin-8, which is a powerful neutrophils chemotactic
factor. Binding of IL-8 to the receptor causes activation of neutrophils. This response is
mediated via a G-protein that activate a phosphatidylinositol-calcium second messenger
system. This receptor binds to IL-8 with a high affinity and to MGSA (GRO) with a low
affinity.
SwissProt: P05362 # ICAM proteins are ligands for the leukocyte adhesion protein LFA-1
(integrin alpha-L/beta-2). During leukocyte trans-endothelial migration, ICAM1 engagement
promotes the assembly of endothelial apical cups through ARHGEF26/SGEF and RHOG
activation. In case of rhinovirus infection acts as a cellular receptor for the virus.
SwissProt: P03956 # Cleaves collagens of types I, II, and III at one site in the helical domain.
Also cleaves collagens of types VII and X. In case of HIV infection, interacts and cleaves the
secreted viral Tat protein, leading to a decrease in neuronal Tat's mediated neurotoxicity.
14
MMP9
matrix
metallopeptidase 9
(gelatinase B, 92kDa
gelatinase, 92kDa type
IV collagenase)
PLAUR
plasminogen activator,
urokinase receptor
SERPINB2
SERPINE1
serpin peptidase
inhibitor, clade B
(ovalbumin), member 2
serpin peptidase
inhibitor, clade E
(nexin, plasminogen
activator inhibitor type
1), member 1
TFPI2
tissue factor pathway
inhibitor 2
TIMP1
TIMP metallopeptidase
inhibitor 1
HIF1A
hypoxia inducible
factor 1, alpha subunit
(basic helix-loop-helix
transcription factor)
SwissProt: P14780 # May play an essential role in local proteolysis of the extracellular matrix
and in leukocyte migration. Could play a role in bone osteoclastic resorption. Cleaves KiSS1
at a Gly- -Leu bond. Cleaves type IV and type V collagen into large C-terminal three quarter
fragments and shorter N-terminal one quarter fragments. Degrades fibronectin but not laminin
or Pz-peptide.
SwissProt: Q03405 # Acts as a receptor for urokinase plasminogen activator. Plays a role in
localizing and promoting plasmin formation. Mediates the proteolysis-independent signal
transduction activation effects of U-PA. It is subject to negative-feedback regulation by U-PA
which cleaves it into an inactive form.
SwissProt: P05120 # Inhibits urokinase-type plasminogen activator. The monocyte derived
PAI-2 is distinct from the endothelial cell-derived PAI-1.
SwissProt: P05121 # Serine protease inhibitor. This inhibitor acts as 'bait' for tissue
plasminogen activator, urokinase, protein C and matriptase-3/TMPRSS7. Its rapid interaction
with PLAT may function as a major control point in the regulation of fibrinolysis.
SwissProt: P48307 # May play a role in the regulation of plasmin-mediated matrix
remodeling. Inhibits trypsin, plasmin, factor VIIa/tissue factor and weakly factor Xa. Has no
effect on thrombin.
SwissProt: P01033 # Complexes with metalloproteinases (such as collagenases) and
irreversibly inactivates them by binding to their catalytic zinc cofactor. Also mediates
erythropoiesis in vitro; but, unlike IL-3, it is species-specific, stimulating the growth and
differentiation of only human and murine erythroid progenitors. Known to act on MMP-1,
MMP-2, MMP-3, MMP-7, MMP-8, MMP-9, MMP-10, MMP-11, MMP-12, MMP-13 and
MMP-16. Does not act on MMP-14.
SwissProt: Q16665 # Functions as a master transcriptional regulator of the adaptive response
to hypoxia. Under hypoxic conditions, activates the transcription of over 40 genes, including
erythropoietin, glucose transporters, glycolytic enzymes, vascular endothelial growth factor,
HILPDA, and other genes whose protein products increase oxygen delivery or facilitate
metabolic adaptation to hypoxia. Plays an essential role in embryonic vascularization, tumor
angiogenesis and pathophysiology of ischemic disease. Binds to core DNA sequence 5'[AG]CGTG-3' within the hypoxia response element (HRE) of target gene promoters.
15
HMOX1
heme oxygenase
(decycling) 1
PRKCZ
protein kinase C, zeta
PTGS2
prostaglandinendoperoxide synthase
2 (prostaglandin G/H
Activation requires recruitment of transcriptional coactivators such as CREBPB and EP300.
Activity is enhanced by interaction with both, NCOA1 or NCOA2. Interaction with redox
regulatory protein APEX seems to activate CTAD and potentiates activation by NCOA1 and
CREBBP. Involved in the axonal distribution and transport of mitochondria in neurons during
hypoxia.
Heme oxygenase, an essential enzyme in heme catabolism, cleaves heme to form biliverdin,
which is subsequently converted to bilirubin by biliverdin reductase, and carbon monoxide, a
putative neurotransmitter. Heme oxygenase activity is induced by its substrate heme and by
various nonheme substances. Heme oxygenase occurs as 2 isozymes, an inducible heme
oxygenase-1 and a constitutive heme oxygenase-2. HMOX1 and HMOX2 belong to the heme
oxygenase family. (provided by RefSeq, Jul 2008)
SwissProt: Q05513 # Calcium- and diacylglycerol-independent serine/threonine-protein
kinase that functions in phosphatidylinositol 3-kinase (PI3K) pathway and mitogen-activated
protein (MAP) kinase cascade, and is involved in NF-kappa-B activation, mitogenic
signaling, cell proliferation, cell polarity, inflammatory response and maintenance of longterm potentiation (LTP). Upon lipopolysaccharide (LPS) treatment in macrophages, or
following mitogenic stimuli, functions downstream of PI3K to activate MAP2K1/MEK1MAPK1/ERK2 signaling cascade independently of RAF1 activation. In EGF-induced cells,
binds and activates MAP2K5/MEK5-MAPK7/ERK5 independently of its kinase activity and
can activate JUN promoter through MEF2C. Through binding with SQSTM1/p62, functions
in interleukin-1 signaling and activation of NF-kappa-B with the specific adapters RIPK1 and
TRAF6. Participates in TNF-dependent transactivation of NF-kappa-B by phosphorylating
and activating IKBKB kinase, which in turn leads to the degradation of NF-kappa-B
inhibitors. In inflammatory response, is required for the T-helper 2 (Th2) differentiation
process, including interleukines production, efficient activation of JAK1 and the subsequent
phosphorylation and nuclear translocation of STAT6. May be involved in development of
allergic airway inflammation (asthma), a process dependent on Th2 immune response. In NFkappa-B-mediated inflammatory response, can relieve the SETD6-dependent repression of
NF-kappa-B target genes by phosphorylating the RELA subunit at 'Ser-311'. Is necessary and
sufficient for LTP maintenance in hippocampal CA1 pyramidal cells.
Prostaglandin-endoperoxide synthase (PTGS), also known as cyclooxygenase, is the key
enzyme in prostaglandin biosynthesis, and acts both as a dioxygenase and as a peroxidase.
There are two isozymes of PTGS: a constitutive PTGS1 and an inducible PTGS2, which
16
synthase and
cyclooxygenase)
SOD2
superoxide dismutase
2, mitochondrial
CD14
CD14 molecule
FOSL1
FOS-like antigen 1
IL10
interleukin 10
IL1B
interleukin 1, beta
IL6
interleukin 6
(interferon, beta 2)
differ in their regulation of expression and tissue distribution. This gene encodes the inducible
isozyme. It is regulated by specific stimulatory events, suggesting that it is responsible for the
prostanoid biosynthesis involved in inflammation and mitogenesis. (provided by RefSeq, Feb
2009)
This gene is a member of the iron/manganese superoxide dismutase family. It encodes a
mitochondrial protein that forms a homotetramer and binds one manganese ion per subunit.
This protein binds to the superoxide byproducts of oxidative phosphorylation and converts
them to hydrogen peroxide and diatomic oxygen. Mutations in this gene have been associated
with idiopathic cardiomyopathy (IDC), premature aging, sporadic motor neuron disease, and
cancer. Alternate transcriptional splice variants, encoding different isoforms, have been
characterized. (provided by RefSeq, Jul 2008)
SwissProt: P08571 # Cooperates with MD-2 and TLR4 to mediate the innate immune
response to bacterial lipopolysaccharide (LPS). Acts via MyD88, TIRAP and TRAF6, leading
to NF-kappa-B activation, cytokine secretion and the inflammatory response. Up-regulates
cell surface molecules, including adhesion molecules
The Fos gene family consists of 4 members: FOS, FOSB, FOSL1, and FOSL2. These genes
encode leucine zipper proteins that can dimerize with proteins of the JUN family, thereby
forming the transcription factor complex AP-1. As such, the FOS proteins have been
implicated as regulators of cell proliferation, differentiation, and transformation. (provided by
RefSeq, Jul 2008)
SwissProt: P22301 # Inhibits the synthesis of a number of cytokines, including IFN-gamma,
IL-2, IL-3, TNF and GM-CSF produced by activated macrophages and by helper T-cells.
SwissProt: P01584 # Produced by activated macrophages, IL-1 stimulates thymocyte
proliferation by inducing IL-2 release, B-cell maturation and proliferation, and fibroblast
growth factor activity. IL-1 proteins are involved in the inflammatory response, being
identified as endogenous pyrogens, and are reported to stimulate the release of prostaglandin
and collagenase from synovial cells.
This gene encodes a cytokine that functions in inflammation and the maturation of B cells. In
addition, the encoded protein has been shown to be an endogenous pyrogen capable of
inducing fever in people with autoimmune diseases or infections. The protein is primarily
produced at sites of acute and chronic inflammation, where it is secreted into the serum and
induces a transcriptional inflammatory response through interleukin 6 receptor, alpha. The
functioning of this gene is implicated in a wide variety of inflammation-associated disease
17
IL8
MEFV
NFKB2
NLRC4
NLRP3
NOD2
states, including suspectibility to diabetes mellitus and systemic juvenile rheumatoid arthritis.
(provided by RefSeq, Jun 2011)
SwissProt: P10145 # IL-8 is a chemotactic factor that attracts neutrophils, basophils, and Tcells, but not monocytes. It is also involved in neutrophil activation. It is released from several
cell types in response to an inflammatory stimulus. IL-8(6-77) has a 5-10-fold higher activity
interleukin 8
on neutrophil activation, IL-8(5-77) has increased activity on neutrophil activation and IL8(7-77) has a higher affinity to receptors CXCR1 and CXCR2 as compared to IL-8(1-77),
respectively.
This gene encodes a protein, also known as pyrin or marenostrin, that is an important
Mediterranean fever
modulator of innate immunity. Mutations in this gene are associated with Mediterranean
fever, a hereditary periodic fever syndrome. (provided by RefSeq, Jul 2008)
This gene encodes one of the subunits of the transcription factor complex nuclear factorkappa-B (NFkB). The NFkB transcription factor complex is expressed in numerous cell types
nuclear factor of kappa and functions as a central activator of genes involved in inflammation and immune function.
light polypeptide gene
The NFkB complex can consist of different subunits that form both homo- or heterodimers
enhancer in B-cells 2
which bind specific kappa-B elements in target genes. This gene encodes the p100 subunit
(p49/p100)
that is processed into the active p52 subunit. This protein can function as both a
transcriptional activator and repressor, depending on its dimer partner. Alternate splicing
results in both coding and non-coding variants. (provided by RefSeq, May 2012)
Uniprot Q9NPP4 # Key component of inflammasomes that indirectly senses specific proteins
NLR family, CARD
from pathogenic bacteria and fungi and responds by assembling an inflammasome complex
domain containing 4
that promotes caspase-1 activation, cytokine production and macrophage pyroptosis.
SwissProt: Q96P20 # May function as an inducer of apoptosis. Interacts selectively with ASC
and this complex may function as an upstream activator of NF-kappa-B signaling. Inhibits
NLR family, pyrin
TNF-alpha induced activation and nuclear translocation of RELA/NF-KB p65. Also inhibits
domain containing 3
transcriptional activity of RELA. Activates caspase-1 in response to a number of triggers
including bacterial or viral infection which leads to processing and release of IL1B and IL18.
This gene is a member of the Nod1/Apaf-1 family and encodes a protein with two caspase
recruitment (CARD) domains and six leucine-rich repeats (LRRs). The protein is primarily
nucleotide-binding
expressed in the peripheral blood leukocytes. It plays a role in the immune response to
oligomerization domain
intracellular bacterial lipopolysaccharides (LPS) by recognizing the muramyl dipeptide
containing 2
(MDP) derived from them and activating the NFKB protein. Mutations in this gene have been
associated with Crohn disease and Blau syndrome. (provided by RefSeq, Jul 2008)
18
TLR2
toll-like receptor 2
TLR4
toll-like receptor 4
TNF
tumor necrosis factor
TREM1
triggering receptor
expressed on myeloid
cells 1
TYROBP
TYRO protein tyrosine
kinase binding protein
SwissProt: O60603 # Cooperates with LY96 to mediate the innate immune response to
bacterial lipoproteins and other microbial cell wall components. Cooperates with TLR1 to
mediate the innate immune response to bacterial lipoproteins or lipopeptides. Acts via
MYD88 and TRAF6, leading to NF-kappa-B activation, cytokine secretion and the
inflammatory response. May also promote apoptosis in response to lipoproteins. Recognizes
mycoplasmal macrophage-activating lipopeptide-2kD (MALP-2), soluble tuberculosis factor
(STF), phenol-soluble modulin (PSM) and B.burgdorferi outer surface protein A lipoprotein
(OspA-L) cooperatively with TLR6.
SwissProt: O00206 # Cooperates with LY96 and CD14 to mediate the innate immune
response to bacterial lipopolysaccharide (LPS). Acts via MYD88, TIRAP and TRAF6,
leading to NF-kappa-B activation, cytokine secretion and the inflammatory response. Also
involved in LPS-independent inflammatory responses triggered by Ni(2+). These responses
require non-conserved histidines and are, therefore, species-specific.
This gene encodes a multifunctional proinflammatory cytokine that belongs to the tumor
necrosis factor (TNF) superfamily. This cytokine is mainly secreted by macrophages. It can
bind to, and thus functions through its receptors TNFRSF1A/TNFR1 and
TNFRSF1B/TNFBR. This cytokine is involved in the regulation of a wide spectrum of
biological processes including cell proliferation, differentiation, apoptosis, lipid metabolism,
and coagulation. This cytokine has been implicated in a variety of diseases, including
autoimmune diseases, insulin resistance, and cancer. Knockout studies in mice also suggested
the neuroprotective function of this cytokine. (provided by RefSeq, Jul 2008)
SwissProt: Q9NP99 # Stimulates neutrophil and monocyte-mediated inflammatory responses.
Triggers release of pro-inflammatory chemokines and cytokines, as well as increased surface
expression of cell activation markers. Amplifier of inflammatory responses that is triggered
by bacterial and fungal infections and is a crucial mediator of septic shock.
This gene encodes a transmembrane signaling polypeptide which contains an immunoreceptor
tyrosine-based activation motif (ITAM) in its cytoplasmic domain. The encoded protein may
associate with the killer-cell inhibitory receptor (KIR) family of membrane glycoproteins and
may act as an activating signal transduction element. This protein may bind zeta-chain (TCR)
associated protein kinase 70kDa (ZAP-70) and spleen tyrosine kinase (SYK) and play a role
in signal transduction, bone modeling, brain myelination, and inflammation. Mutations within
this gene have been associated with polycystic lipomembranous osteodysplasia with
sclerosing leukoencephalopathy (PLOSL), also known as Nasu-Hakola disease. Its putative
19
CASQ2
calsequestrin 2 (cardiac
muscle)
GALR2
galanin receptor 2
GNA11
guanine nucleotide
binding protein (G
protein), alpha 11 (Gq
class)
KCNMA1
potassium large
conductance calciumactivated channel,
subfamily M, alpha
member 1
MYH11
myosin, heavy chain
11, smooth muscle
receptor, triggering receptor expressed on myeloid cells 2 (TREM2), also causes PLOSL.
Multiple alternative transcript variants encoding distinct isoforms have been identified for this
gene. (provided by RefSeq, Mar 2010)
SwissProt: O14958 # Calsequestrin is a high-capacity, moderate affinity, calcium-binding
protein and thus acts as an internal calcium store in muscle. The release of calcium bound to
calsequestrin through a calcium release channel triggers muscle contraction. The skeletal
muscle isoform (CASQ1) binds around 80 Ca(2+) ions, while the cardiac isoform (CASQ2)
binds approximately 60 Ca(2+) ions.
Galanin is an important neuromodulator present in the brain, gastrointestinal system, and
hypothalamopituitary axis. It is a 30-amino acid non-C-terminally amidated peptide that
potently stimulates growth hormone secretion, inhibits cardiac vagal slowing of heart rate,
abolishes sinus arrhythmia, and inhibits postprandial gastrointestinal motility. The actions of
galanin are mediated through interaction with specific membrane receptors that are members
of the 7-transmembrane family of G protein-coupled receptors. GALR2 interacts with the Nterminal residues of the galanin peptide. The primary signaling mechanism for GALR2 is
through the phospholipase C/protein kinase C pathway (via Gq), in contrast to GALR1, which
communicates its intracellular signal by inhibition of adenylyl cyclase through Gi. However,
it has been demonstrated that GALR2 couples efficiently to both the Gq and Gi proteins to
simultaneously activate 2 independent signal transduction pathways. (provided by RefSeq, Jul
2008)
SwissProt: P29992 # Guanine nucleotide-binding proteins (G proteins) are involved as
modulators or transducers in various transmembrane signaling systems. Acts as an activator of
phospholipase C.
MaxiK channels are large conductance, voltage and calcium-sensitive potassium channels
which are fundamental to thecontrol of smooth muscle tone and neuronal excitability. MaxiK
channels can be formed by 2 subunits: the pore-forming alpha subunit, which is the product of
this gene, and the modulatory beta subunit. Intracellular calcium regulates the physical
association between the alpha and beta subunits. Alternatively spliced transcript variants
encoding different isoforms have been identified. (provided by RefSeq, Jul 2008)
The protein encoded by this gene is a smooth muscle myosin belonging to the myosin heavy
chain family. The gene product is a subunit of a hexameric protein that consists of two heavy
chain subunits and two pairs of non-identical light chain subunits. It functions as a major
20
MYLK
myosin light chain
kinase
contractile protein, converting chemical energy into mechanical energy through the hydrolysis
of ATP. The gene encoding a human ortholog of rat NUDE1 is transcribed from the reverse
strand of this gene, and its 3' end overlaps with that of the latter. The pericentric inversion of
chromosome 16 (inv(16)(p13q22)) produces a chimeric transcript that encodes a protein
consisting of the first 165 residues from the N terminus of core-binding factor beta in a fusion
with the C-terminal portion of the smooth muscle myosin heavy chain. This chromosomal
rearrangement is associated with acute myeloid leukemia of the M4Eo subtype. Alternative
splicing generates isoforms that are differentially expressed, with ratios changing during
muscle cell maturation. Alternatively spliced transcript variants encoding different isoforms
have been identified. (provided by RefSeq, Jul 2008)
SwissProt: Q15746 # Calcium/calmodulin-dependent myosin light chain kinase implicated in
smooth muscle contraction via phosphorylation of myosin light chains (MLC). Also regulates
actin-myosin interaction through a non-kinase activty. Phosphorylates PTK2B/PYK2 and
myosin light-chains. Involved in the inflammatory response (e.g. apoptosis, vascular
permeability, leukocyte diapedesis), cell motility and morphology, airway hyperreactivity and
other activities relevant to asthma. Required for tonic airway smooth muscle contraction that
is necessary for physiological and asthmatic airway resistance. Necessary for gastrointestinal
motility. Implicated in the regulation of endothelial as well as vascular permeability, probably
via the regulation of cytoskeletal rearrangements. In the nervous system it has been shown to
control the growth initiation of astrocytic processes in culture and to participate in transmitter
release at synapses formed between cultured sympathetic ganglion cells. Critical participant in
signaling sequences that result in fibroblast apoptosis. Plays a role in the regulation of
epithelial cell survival. Required for epithelial wound healing, especially during actomyosin
ring contraction during purse-string wound closure. Mediates RhoA-dependent membrane
blebbing. Triggers TRPC5 channel activity in a calcium-dependent signaling, by inducing its
subcellular localization at the plasma membrane. Promotes cell migration (including tumor
cells) and tumor metastasis. PTK2B/PYK2 activation by phosphorylation mediates ITGB2
activation and is thus essential to trigger neutrophil transmigration during acute lung injury
(ALI). May regulate optic nerve head astrocyte migration. Probably involved in mitotic
cytoskeletal regulation. Regulates tight junction probably by modulating ZO-1 exchange in
the perijunctional actomyosin ring. Mediates burn-induced microvascular barrier injury;
triggers endothelial contraction in the development of microvascular hyperpermeability by
phosphorylating MLC. Essential for intestinal barrier dysfunction. Mediates Giardia spp.-
21
MYOM1
myomesin family
member 1
Information
retrieved
from
mediated reduced epithelial barrier function during giardiasis intestinal infection via
reorganization of cytoskeletal F-actin and tight junctional ZO-1.
The giant protein titin, together with its associated proteins, interconnects the major structure
of sarcomeres, the M bands and Z discs. The C-terminal end of the titin string extends into the
M line, where it binds tightly to M-band constituents of apparent molecular masses of 190 kD
(myomesin 1) and 165 kD (myomesin 2). This protein, myomesin 1, like myomesin 2, titin,
and other myofibrillar proteins contains structural modules with strong homology to either
fibronectin type III (motif I) or immunoglobulin C2 (motif II) domains. (provided by RefSeq,
Jul 2008)
the
gene
summary
section
of
GeneCards
webpage
contributed
by
(http://www.ncbi.nlm.nih.gov/gene) and UniProtKB (UniprotKB/Swiss-Prot/UniprotKB/TrEMBL)*.
*Reorganizing the protein space at the Universal Protein Resource (UniProt). Nucleic Acids Res 2012;40:D71-D75.
22
Entrez
Gene
Supplemental Table 5. Comparison of target gene expressions between fetal-CTL (n=4)
and Surg-CTL (n=14) by qPCR assay
Surg-CTL vs. Fetal-CTL
Fold
P-value
Gene
Angiogensis
ACE
ACE2
ANGPLT4
FLT1
GJA5
HBEGF
NR1H4
THBS1
0.678
0.925
1.003
0.309
0.745
1.664
1.408
1.011
0.288
1.000
0.750
0.008
0.203
0.671
0.457
1.000
Arginine Metabolism
ARG1
CPS1
OAT
OTC
1.223
0.799
0.876
0.615
1.000
0.671
0.915
0.339
Cell Adhesion and Chemotaxis
CCL20
CD44
CXCL5
CXCR1
ICAM1
3.054
0.798
1.786
2.215
0.765
0.288
0.243
0.832
0.111
0.111
ECM Remodeling
MMP1
MMP9
PLAUR
SERPINB2
SERPINE1
TFPI2
TIMP1
0.442
0.337
1.005
2.319
1.194
0.558
0.877
0.671
0.167
0.457
0.111
0.915
0.089
0.366
Hypoxia and Oxidative Stress
HIF1A
HMOX1
0.827
0.727
0.457
0.457
23
PRKCZ
PTGS2
SOD2
0.474
2.589
0.884
0.012
0.056
0.457
Inflammation
CD14
FOSL1
IL10
IL1B
IL6
IL8
MEFV
NFKB2
NLRC4
NLRP3
NOD2
TLR2
TLR4
TNF
TREM1
TYROBP
1.494
6.028
1.480
2.513
13.549
6.423
1.158
0.465
1.681
1.512
0.638
1.188
1.106
1.898
3.278
1.265
0.089
1.000
0.202
0.089
0.015
0.026
0.832
0.008
0.089
0.288
0.071
0.457
0.524
0.089
0.056
0.457
Muscle Contraction
CASQ2
GALR2
GNA11
KCNMA1
MYH11
MYLK
MYOM1
1.403
1.434
0.815
1.100
1.345
1.255
0.674
0.750
0.595
0.243
0.832
0.832
0.750
0.288
24
Supplemental Table 6. Comparison of target gene expressions in small intestinal tissues of
NEC (n=12), SIP (n=11) and Surg-CTL (n=9) preterm infants by qPCR assay
Gene
Angiogensis
ACE
ACE2
ANGPTL4
FLT1
GJA5
HBEGF
NR1H4
THBS1
NEC vs. Surg-CTL
Fold
P-value
SIP vs. Surg-CTL
Fold
P-value
NEC vs. SIP
Fold
P-value
0.060
0.079
9.673
1.799
1.680
1.594
0.116
3.367
0.002
0.011
0.021
0.034
0.063
0.483
0.016
0.004
0.496
0.748
4.624
1.034
1.101
1.028
0.790
2.109
0.124
0.739
0.097
0.994
0.923
1.000
0.836
0.178
0.121
0.106
2.092
1.740
1.525
1.551
0.147
1.597
0.052
0.003
0.294
0.045
0.169
0.367
0.003
0.254
83.346
0.088
0.086
0.052
0.007
0.000
0.000
0.001
10.936
1.090
0.665
4.122
0.086
0.952
0.405
0.539
7.621
0.081
0.129
0.013
0.015
0.000
0.030
0.334
Cell Adhesion and Chemotaxis
CCL20
12.543
CD44
1.558
CXCL5
246.244
CXCR1
5.078
ICAM1
2.921
0.004
0.055
0.007
0.011
0.028
1.001
0.577
11.461
1.957
1.056
1.000
0.105
0.476
0.248
0.998
12.536
2.699
21.485
2.594
2.767
0.004
0.000
0.009
0.054
0.027
ECM Remodeling
MMP1
MMP9
PLAUR
SERPINB2
SERPINE1
TFPI2
TIMP1
0.003
0.020
0.000
0.038
0.034
0.041
0.001
8.686
1.197
2.368
28.483
3.651
11.636
4.666
0.040
0.973
0.248
0.073
0.120
0.013
0.002
2.266
4.115
6.468
48.568
3.135
4.431
3.026
0.105
0.025
0.001
0.042
0.139
0.120
0.008
0.000
0.006
1.175
2.729
0.735
0.048
1.961
1.398
0.003
0.575
Arginine Metabolism
ARG1
CPS1
OAT
OTC
19.682
4.926
15.320
1383.367
11.448
51.564
14.119
Hypoxia and Oxidative Stress
HIF1A
2.305
HMOX1
3.817
25
PRKCZ
PTGS2
SOD2
0.160
11.194
11.400
0.002
0.026
0.000
0.749
3.473
2.125
0.520
0.099
0.102
0.214
3.223
5.363
0.001
0.108
0.000
Inflammation
CD14
FOSL1
IL10
IL1B
IL6
IL8
MEFV
NFKB2
NLRC4
NLRP3
NOD2
TLR2
TLR4
TNF
TREM1
TYROBP
5.510
5.388
4.308
71.685
20.474
240.474
5.129
1.677
5.275
3.716
5.258
6.953
1.686
7.068
22.595
2.240
0.009
0.025
0.032
0.003
0.009
0.003
0.023
0.025
0.002
0.018
0.002
0.001
0.150
0.018
0.000
0.042
1.754
1.645
1.331
5.205
2.238
12.780
2.022
0.910
1.525
1.122
1.385
1.682
0.950
1.107
3.512
0.969
0.146
0.799
0.824
0.344
0.493
0.151
0.070
0.954
0.287
0.975
0.408
0.412
0.983
0.977
0.063
0.997
3.142
3.274
3.237
13.771
9.147
18.817
2.537
1.842
3.459
3.312
3.797
4.133
1.775
6.387
6.435
2.311
0.028
0.053
0.062
0.004
0.014
0.004
0.100
0.005
0.006
0.024
0.004
0.002
0.106
0.020
0.001
0.036
0.139
0.479
0.222
0.279
0.181
0.343
0.204
0.012
0.194
0.000
0.057
0.017
0.017
0.002
0.064
0.210
0.600
0.154
0.138
0.308
0.185
0.007
0.004
0.030
0.022
0.013
0.011
0.002
2.151
2.278
0.370
1.809
1.314
1.116
1.103
0.365
0.555
0.029
0.710
0.813
0.982
0.984
Muscle Contraction
CASQ2
GALR2
GNA11
KCNMA1
MYH11
MYLK
MYOM1
26