SUPPLEMENTAL DATA
1. Supplemental methods
1.1 Echocardiography
Vigil mice were studied on an echocardiography system (Vivid7 General Electrics; 13 MHz linear
transducer). Cardiac dimensions and function were assessed by M-mode and 2-dimensional
echocardiography. Left ventricle (LV) end-diastolic and end-systolic volumes were measured with the arealength method. Heart rate was determined by using pulse wave Doppler interrogation of mitral inflow.
Cardiac output and LV ejection fraction based on these parameters were calculated to access cardiac
function.
1.2 Surgical preparation and experimental protocol
Investigation conforms to the Directive 2010/63/EU of the European Parliament. The surgical procedures
were approved by the regional Committee for animal research (authorization number CE-LR-0814).
Acute myocardial ischemia and reperfusion were performed in C57BL/6 (Charles River Laboratoire,
France), Zac1-/- knockout and Zac1+/+ wild-type (WT) littermates mice (L. Journot, IGF Montpellier). Male
mice (22-30 g) were anaesthetized with an intramuscular (IM) injection of an anesthetic mixture comprising
ketamine (50 mg/kg; Imalgène® 500; Merial), xylazine (10 mg/kg; Rompun® 2%; Bayer) and
chlorpromazine (1.25 mg/kg; Largactil® 5 mg/ml; Sanofi-Aventis). Mice were ventilated via a tracheal
intubation on a Harvard rodent respirator. The body temperature was maintained constant (36.8°C-37°C).
After a second injection of ketamine (50 mg/kg) and xylazine (10 mg/kg), the chest was opened by a left
lateral thoracotomy and a reversible coronary artery snare occluder was placed around the left coronary
artery. 1 The adequacy of anesthesia and pain analgesia was monitored during the surgical protocol by
pinching the end of the posterior legs. All mice underwent 40 min of ischemia followed by 60 min of
reperfusion by loosening the knot and were randomly assigned to the different groups (Figure 1A):
- The Ischemia/Reperfusion group (IR) underwent 40 minutes of ischemia and 60 minutes of reperfusion.
- The preconditioning group (PreC) underwent the PreC algorithm, 3 cycles of 1 minute -reperfusion and 1
minute - reocclusion, before the onset of ischemia.
- The postconditioning group (PostC) underwent a protocol comprising, after the 40 minutes of ischemia, 3
cycles of 1 minute - reperfusion and 1 minute - reocclusion preceeding the 60 minutes of reperfusion.
After reperfusion, the knot is finally tightened, and a phtalocyanin blue dye solution injected in the left
ventricular cavity. The heart is harvested, washed in a 4°C-cold PBS buffer, the right ventricle and atrias
removed.
1.3 Infarct size assessment
The left ventricle was sliced transversally into 1 mm thick-sections and incubated in a 1% solution of 2,3,5triphenyltetrazolium chloride (TTC; Sigma Aldrich) for 15 minutes at 37°C. After fixation in a 4%
paraformaldehyde-phosphate buffer saline (PFA-PBS) solution, the slices were weighed and each side was
photographed with an Olympus camera. The ischemic risk area (unstained in blue) and the infarcted area
(unstained by TTC) were measured by planimetry using Image J (Scion corp., Frederick, MD). Infarct size
was expressed as a percentage of the ischemic risk area.
1.4 DNA fragmentation Assay
Specific DNA fragmentation was quantified in transmural samples of nonischemic or ischemic areas of the
LV with an enzyme-linked immunosorbent assay kit (Roche Diagnostics) designed to measure the amount
of cytosolic oligonucleosome-bound DNA, as previously described.2 Transmural samples from 30 mg of
non-ischemic areas of the left ventricle (central portion of the septum) and ischemic areas (portion of the LV
free wall directly under the silk ligation) were harvested on mice submitted to the surgical protocol. Tissues
samples were disintegrated in a tissue grinder in 400 µl of lysis buffer supplied with the kit. The homogenate
was centrifuged at 13,000 g for 10 min. The supernatant was used as antigen source in the sandwich ELISA.
Incubation buffer, instead of the sample solution, and DNA-histone complex were used as negative and
positive controls, respectively. Two values from the double absorbance measurements (405 nm / 490 nm) of
the samples were averaged, and the background (negative control) was subtracted from each of these
averages. The positive control was used as an internal control for daily variability. DNA soluble nucleosome
was quantified both in blue-colored region (non-ischemic region = NI) as well as in the non blue-colored
areas (ischemic region = I). This allows to normalize the rate of DNA fragmentation (internal control) and to
avoid the introduction of variability in the results due to differences in temperature, timing of the
measurement.
1.5 Microarray experiments
RNA extraction
At the end of surgery, the left ventricular tissue was rinsed using cold PBS (4°C). Non blue (ischemic)
regions of left ventricles were microdissected and incubated in RNA later (1/5: vol/vol; Ambion) overnight
at 4°C and stored at -80°C. Total RNAs were extracted using Trizol LS reagent (Invitrogen, Basel,
Switzerland) according to the manufacturer’s instructions. RNA precipitation was obtained using the
chloroform-isopropanol method. Pellets were washed with 70% ethanol and eluted in RNase-free water.
RNA samples extracted from different mice subjected to IR (n=9), PreC (n=9) or PostC (n=9) were subpooled in 3 mixtures (IR1,2,3; PreC1,2,3; PostC1,2,3).
RNA labelling
For the preparation of the labelled Cy3- and Cy5- aRNA target, one microgram of total RNA samples were
amplified and synthesized using Amino Allyl MessageAmpTM II aRNA Amplification kit (Ambion),
according to the manufacturer’s instructions. One-half of the RNA derived from each condition (IR, Prec,
PostC) was labelled with Cy3 and the other half with Cy5 to account for any bias in dye coupling or
emission efficiency.
Microarray plateform
Mouse oligo microarrays were used for gene expression profiling (Montpellier GenomiX Facilities). The
microarrays were designed with the Mouse Array-Ready Oligo Set™ Version 2 (Qiagen Operon)
(http://omad.operon.com/download/index.php). The Mouse oligo-chips contain 16,423 oligonucleotides (70mers) probes representing 16,463 unique genes. All probes were designed from representative sequences of
the UniGene Database Build Mm 102 (February 2002) and the Mouse Reference Sequence (RefSeq)
Database. The library also included sets of positive and negative controls that were used for quality control
purposes.
Hybridization
Prior to hybridization, excess oligonucleotides were removed from the arrays by shaking them twice 1min in
0.2% SDS. Arrays were then washed twice with distilled water.
The two labelled aRNA were added to a microarray hybridization buffer version 2 (GE Healthcare) in a final
concentration of 50% formamide, then denaturated at 95°C for 3 min and applied to the microarrays in
individual chambers of an automated slide processor (GE Healthcare). Hybridization was carried out at 37°C
for 12 h. Hybridized slides were washed at 37°C successively with 1X SSC plus 0.2% SDS for 10 min,
twice with 0.1X SSC plus 0.2% SDS for 10 min, with 0.1X SSC for 1 min and with isopropanol before airdrying.
Data acquisition
Microarrays were immediately scanned in both Cy3 and Cy5 channels with GenePix 4200AL scanner
(Molecular Devices) at 10 µm resolution with a variable PMT voltage to obtain maximal signal intensities
with less than 0.1% probe saturation. ArrayVision software was used for feature extraction. Spots with high
local background or contamination fluorescence were flagged manually. Local background noise was
calculated for each spot as the median values of the fluorescence intensities of 4 squares surrounding the
spot.
Normalization and computation of expression values
No spatial bias in the quality analysis has been detected. A Loess normalisation was performed for all
microarrays to correct technical bias and dye effects. No background correction was performed. Tests of
differential expression were conducted using the LIMMA3 and the Siggenes packages from Bioconductor.
Multiple testing adjustments were performed using a false discovery rate approach.4
These two analyses provide ranking of significantly expressed genes (P<0.001). The gene lists obtained by
LIMMA were used to generate Venn diagrams. The Bioarray Software Environment (BASE,
http://baseprod.igf.cnrs.fr/index.phtml) was used to visualize differential expression for each gene.5
Transcriptomic analysis
cDNA pangenomic microarrays were used to characterize and compare genomic responses to PreC and
PostC. Gene expression profiles were obtained for hearts subjected to IR protocols in both ischemic and non
ischemic parts, and for the ischemic part from hearts subjected to PreC or PostC treatments.
Gene Ontology
Data were analyzed through the use of Ingenuity Pathways Analysis (IPA) 8.7 (Ingenuity® Systems,
www.ingenuity.com).
The Functional Analysis identified the biological functions that were most significant to the data set.
Molecules from the dataset identified as significantly differentially expressed with the LIMMA test were
associated with biological functions in Ingenuity’s Knowledge Base and considered for the analysis. Righttailed Fisher’s exact test was used to calculate a p-value determining the probability that each biological
function assigned to that data set is due to chance alone.
1.6 Quantification of mRNA expression levels by real-time PCR
RNA (1 μg) was first reverse transcribed using Superscript III reverse transcriptase (Invitrogen) and 250 ng
of random hexamer (Amersham Biosciences Europe, Orsay, France) in a final volume of 20 μl. Real-time
PCR analyses of genes and housekeeping genes were performed using SYBR Green PCR master mix
(Applied Biosystems, Foster City, CA) with 1:10 of the reverse-transcription reaction, and were carried out
on an ABI 7500 Sequence Detector (Applied Biosystems). Table S1 summarizes primer sequences
(Supplementary material online).
1.7 Quantification of mRNA expression levels by real-time PCR
RNA (1 μg) was first reverse transcribed using Superscript III reverse transcriptase (Invitrogen) and 250 ng
of random hexamer (Amersham Biosciences Europe, Orsay, France) in a final volume of 20 μl. Real-time
PCR analyses of genes and housekeeping genes were performed using SYBR Green PCR master mix
(Applied Biosystems, Foster City, CA) with 1:10 of the reverse-transcription reaction, and were carried out
on an ABI 7500 Sequence Detector (Applied Biosystems). Primer sequences are given in Table S1. (See
Supplemental for complementary informations). The concentration of the primers used was 300 nM. After
an initial denaturation step for 10 min at 95°C, the thermal cycling conditions were 40 cycles at 95°C for 15
s and 60°C for 1 min. Each sample value was determined from triplicate measurements. The selection of
appropriate housekeeping genes was performed with geNorm (Vandesompele et al., 2002). Expression of
transcripts was normalized to the geometric mean of the expression levels of three housekeeping genes, Hprt
(hypoxanthine-guanine phosphoribosyltransferase), Trfr (Transferrine Receptor) and β actin, according to
the formula Cx/geometric mean (R1, R2, R3) = 2-(Ct[Cx]-arithmetic mean [Ct(R1),Ct(R2),Ct(R3)]), where Ct is the
threshold cycle, and R1, R2, R3 are the three reference genes.
1.8 Immunostaining
At the end of surgery, left ventricles were fixated in 4%-PFA and embedded in paraffin. Paraffin-embedded
LV sections (4 µm) were deparaffinised then rehydrated through an alcohol gradient and washed in
phosphate-buffered saline (PBS). Antigen retrieval was achieved using proteinase K treatment (Fungus,
Invitrogen; 20 µg/ml, 15 minutes). Tissue were washed thoroughly in PBS, and blocked with 3% bovine
serum albumin (BSA, Sigma-Aldrich) and mouse IgG (1:200, Paris Biotech, France). Sections were
incubated 18h at room temperature (RT) with a rabbit ZAC1 antibody (homemade, L. Journot; 1:500) and a
mouse α-actinin antibody (A7732, 1:100; Sigma-Aldrich) at 4°C overnight. After primary antibody
incubation, the sections were washed in PBS, and then incubated (3 hours at RT) with secondary antibody
(Cy3-conjugated anti-rabbit and Alexa488-conjugated anti-mouse, 1:2,000, Jackson ImmunoRes
Laboratories, Inc., West Grove, PA). Primary and secondary antibodies were diluted in PBS containing 3%
BSA and 0.1% Triton X100. Cell nuclei were stained with DAPI (Sigma-Aldrich). After rinsing, stained
sections were mounted in PVA-DABCO (Sigma-Aldrich) and imaged with a Leica DM6000-1. For each LV
section, ZAC1-positive nuclei were randomly counted on at least 10 independent fields. Adobe Photoshop
(Adobe Systems, Paris, France) was used to prepare final figures.
1.9 Statistical analysis
Values are expressed as mean  SDs for in vivo experiments and as mean  SEMs for QPCR experiments.
Multiple comparisons between groups were assessed by the Kruskal-Wallis non parametric test followed by
the Dunn's post hoc test when appropriate. Mann-Whitney comparison was used when only two groups are
compared. Probability values <0.05 were accepted as statistically significant, and the probability values were
noted as follows: P=ns for P>0.05; *P<0.05; **P<0.01; ***P<0.001.
For immunohistochemistry counting, 2x3 contingency table was used for testing the proportion of ZAC1
positive nuclei, followed by a Complete Tukey's HSD multiple comparisons test among proportions.6 All
data were analyzed using GraphPad Prism (GraphPad Software, San Diego California USA) and Matlab
(The Mathworks Inc., Natick MA USA).
References
1.
Roubille F, Franck-Miclo A, Covinhes A, Lafont C, Cransac F, Combes S, et al. Delayed
postconditioning in the mouse heart in vivo. Circulation 2011;124:1330-1336.
2.
Piot CA, Padmanaban D, Ursell PC, Sievers RE, Wolfe CL. Ischemic preconditioning decreases
apoptosis in rat hearts in vivo. Circulation 1997;96:1598-1604.
3.
Smyth GK. Linear models and empirical bayes methods for assessing differential expression in
microarray experiments. Stat Appl Genet Mol Biol 2004;3:Article3.
4.
Reiner A, Yekutieli D, Benjamini Y. Identifying differentially expressed genes using false discovery
rate controlling procedures. Bioinformatics 2003;19:368-375.
5.
Saal LH, Troein C, Vallon-Christersson J, Gruvberger S, Borg A, Peterson C. BioArray Software
Environment (BASE): a platform for comprehensive management and analysis of microarray data.
Genome Biol 2002;3:SOFTWARE0003.
6.
Zar JH. Biostatistical analysis. Upper Saddle River, NJ, USA Prentice-Hall, Inc., 1999.
2. Figure Legends
Figure S1: Validation by quantitative real-time RT-PCR
(A): PostC/IR and PreC/IR ratios obtained with both microarray and quantitative real-time RT-PCR
experiments for 15 cardiovascular-related genes were compared. Data were obtained using the same pooled
RNA extracts used for microarray and real-time RT-PCR (from n=8 IR, n=8 PreC and n=8 PostC mice).
(B): Spearman's correlation coefficient was 0.736.
3. Figures /Tables
Table S1: Primers used for quantitative real time RT-PCR. The accession number is the identifier in the
NCBI nucleotide database (http://www.ncbi.nlm.nih.gov/nucleotide).
Gene Symbol Accession Number Forward sequence (5'-3')
Reverse sequence (3'-5')
Ankrd1
Bag1
Bclaf1
Daxx
Dpf2
Fgf1
Hccs
Ifit3
Lgals3bp
Ltbp1
Lum
Mif
Sgms1
Stk39
Twsg1
Zac1
HKGs:
Actb
Hprt
Trfr
NM_013468.3
GGT GAAT GGT GT CT AAT T GC
GT T ACT GCGT GCCT T CT G
NM_009736.2
AAGAAAGT GACCCGT AGC
T GCCACAGT T ACT T CCT C
CAACT CT T AACT T GCCT CCCT GT T
GT GGGT T T GAGAAAAGCT AACGT T
NM_007829.4
CT T CCGGT CT GT T GT GGGGT CT
AACGGT CT CT CGAGGAT CCCCT
NM_011262.3
GGCT ACCCAGACAT T T CC
GCACAT T AT CCAACT T CT CC
NM_010197.3
T CCT CGGACT CACT AT GG
AACACT CAGAACAGACT CC
NM_008222.3
GCAACAT CT GACCT GACC
GGCAT T CCACAT AAT CAT AGG
NM_010501.1
CACAGCT GAAGT GCCAT T T CA
T T GCACACCCT GT CT T CCAT AT
NM_011150.2
ACCT GGAGGGCACAAAGAT AT G
T GGT AGCT AT T GT ACCGGGCT ACT
NM_206958.1
AGACCCAGACAGT CCAT T CT ACGT
T AGCAGCCACGGGAT ACACAT
NM_008524.2
GCCAAT ACT ACGAT T AT GACAT C
T GCCAGGAGGAACCAT T G
NM_010798,2
GGT CT ACAT CAACT AT T ACG
AT AAACACAGAACACT ACG
NM_144792.3
AT GAT AGAGACCCT GAAGAT G
GAAT GT CAACGCCAAT GC
NM_016866.2
AGAGGCT AT GACT T CAAGGCT GAT AT
CGCT GCT CCGGT T GCT
NM_023053.2
AAT GCAAACCAAAGCAGT AAGT CAT
CT CCT AGCAT ACCAGAACCAAGAT T
NM_009538.1
T CT CCAAGT AT AAGCT GAT GAGACACAT
ACACGCGT AGGAGAT CT T GT T G
NM_007393.3
GCAT T GT GAT GGACT CCGGT
CGT AGAT GGGCACAGT GT GG
NM_013556
GCAGT ACAGCCCCAAAAT GG
GGT CCT T T T CACCAGCAAGCT
AGACCT T GCACT CT T T GGACAT G
GGT GT GT AT GGAT CACCAGT T CCT A
NM_001025393.1
NM_011638.4
Table 1: Primers used for quantitative real time RT-PCR. The Accession Number is the identifier in the NCBI
Table S2: PreC gene list obtained by LIMMA.
Notes: (*) for (includes EG:10216);
Bold: genes jointly regulated by both PreC and PostC.
Fold
change
-4,423
-1,933
-1,668
-1,623
-1,561
-1,515
-1,495
-1,457
-1,417
-1,336
-1,316
-1,316
-1,309
-1,303
-1,287
-1,28
-1,273
-1,272
-1,269
-1,251
-1,249
-1,241
-1,237
-1,236
-1,234
-1,208
-1,192
-1,19
-1,188
-1,184
-1,183
1,129
1,191
1,306
1,329
1,409
1,41
1,472
1,798
ID
Symbol
Hspa1a
Anp32a
Adamts1
Prg4
C3
Timm44
Phlda1
Trh
Emd
Plagl1
Ppp1r10
Atp1a1
Timp1
Fasn
Jmjd1c
Ehd4
Flnc
Palld
Pmfbp1
Tsc22d3
Cgnl1
Tmx4
Csrp2
Degs2
Brd2
Ltbp1
Trpc2
Med25
Solh
Bclaf1
Twsg1
Stk39
Lrrc2
Lgals3bp
Tmem140
Bmp5
Ifit3
Zfp287
Ptgds
HSPA1A
ANP32A
ADAMTS1
PRG4 (*)
NAD+
TIMM44
PHLDA1
TRH
EMD
PLAGL1
PPP1R10
ATP1A1
TIMP1
FASN
JMJD1C
EHD4
FLNC
PALLD
PMFBP1
TSC22D3
CGNL1
TMX4
CSRP2
DEGS2
BRD2
LTBP1
TRPC2
MED25
SOLH
BCLAF1
TWSG1
STK39
LRRC2
LGALS3BP
TMEM140
BMP5
IFIT3
PTGDS
Gene Name
heat shock 70kDa protein 1A
acidic nuclear phosphoprotein 32 family, member A
ADAM metallopeptidase with thrombospondin type1 motif, 1
proteoglycan 4
translocase of inner mitochondrial membrane 44 homolog (yeast)
pleckstrin homology-like domain, family A, member 1
thyrotropin-releasing hormone
emerin
pleiomorphic adenoma gene-like 1
protein phosphatase 1, regulatory (inhibitor) subunit 10
ATPase, Na+/K+ transporting, alpha 1 polypeptide
TIMP metallopeptidase inhibitor 1
fatty acid synthase
jumonji domain containing 1C
EH-domain containing 4
filamin C, gamma
palladin, cytoskeletal associated protein
polyamine modulated factor 1 binding protein 1
TSC22 domain family, member 3
cingulin-like 1
thioredoxin-related transmembrane protein 4
cysteine and glycine-rich protein 2
degenerative spermatocyte homolog 2, lipid desaturase (Drosophila)
bromodomain containing 2
latent transforming growth factor beta binding protein 1
transient receptor potential cation channel, subfamily C, member 2, pseudogene
mediator complex subunit 25
small optic lobes homolog (Drosophila)
BCL2-associated transcription factor 1
twisted gastrulation homolog 1 (Drosophila)
serine threonine kinase 39 (STE20/SPS1 homolog, yeast)
leucine rich repeat containing 2
lectin, galactoside-binding, soluble, 3 binding protein
transmembrane protein 140
bone morphogenetic protein 5
interferon-induced protein with tetratricopeptide repeats 3
prostaglandin D2 synthase 21kDa (brain)
Table S3: PostC gene list obtained by LIMMA.
Notes: * for includes EG:10216; †: includes EG:2597); ‡: includes EG:7323; §: includes EG:5160); ||:
includes EG:10728); #: includes EG:25797); **: includes EG:54946);
Bold: genes jointly regulated by both PreC and PostC.
Fold
change
-2,148
-1,812
-1,761
-1,651
-1,645
-1,624
-1,617
-1,608
-1,597
-1,575
-1,564
-1,541
-1,537
-1,531
-1,527
-1,52
-1,506
-1,502
-1,501
-1,484
-1,481
-1,476
-1,461
-1,452
-1,449
-1,449
-1,447
-1,444
-1,439
-1,435
-1,428
-1,428
-1,427
-1,412
-1,406
-1,392
-1,379
-1,376
-1,372
-1,372
-1,371
-1,37
-1,364
-1,361
-1,359
-1,358
-1,356
-1,355
-1,354
ID
Symbol
Gene name
Casq1
Smyd1
Ankrd1
Npas3
Ppp3r1
Pitpnb
Utrn
Clec14a
Fuca2
Tgfbr3
Ctgf
F2r
Fgl2
Trim35
Lum
Opa1
Prg4
Nfe2l2
Dgat2
Slc25a33
Tmem38a
Rab18
Mlf1
Emp1
Mapkapk2
Nat13
Peli1
Ahcyl1
Ifngr2
Hnrnpk
Sfrs1
Arhgdia
Fstl1
Golph3
Sgms1
Gapdh
Eif2s3x
Hnrnpa3
Sypl
Mlana
Dnaja3
Col5a2
Ube2g1
Chmp4c
Hpgd
Obfc2a
Garnl1
Atp1a2
Galnt1
CASQ1
SMYD1
ANKRD1
NPAS3
PPP3R1
PITPNB
UTRN
CLEC14A
FUCA2
TGFBR3
CTGF
F2R
FGL2
TRIM35
LUM
OPA1
PRG4 *
NFE2L2
DGAT2
SLC25A33
TMEM38A
RAB18
MLF1
EMP1
MAPKAPK2
NAA50
PELI1
AHCYL1
IFNGR2
HNRNPK
SFRS1
ARHGDIA
FSTL1
GOLPH3
SGMS1
GAPDH †
calsequestrin 1 (fast-twitch, skeletal muscle)
SET and MYND domain containing 1
ankyrin repeat domain 1 (cardiac muscle)
neuronal PAS domain protein 3
protein phosphatase 3, regulatory subunit B, alpha
phosphatidylinositol transfer protein, beta
utrophin
C-type lectin domain family 14, member A
fucosidase, alpha-L- 2, plasma
transforming growth factor, beta receptor III
connective tissue growth factor
coagulation factor II (thrombin) receptor
fibrinogen-like 2
tripartite motif-containing 35
lumican
optic atrophy 1 (autosomal dominant)
proteoglycan 4
nuclear factor (erythroid-derived 2)-like 2
diacylglycerol O-acyltransferase homolog 2 (mouse)
solute carrier family 25, member 33
transmembrane protein 38A
RAB18, member RAS oncogene family
myeloid leukemia factor 1
epithelial membrane protein 1
mitogen-activated protein kinase-activated protein kinase 2
N(alpha)-acetyltransferase 50, NatE catalytic subunit
pellino homolog 1 (Drosophila)
adenosylhomocysteinase-like 1
interferon gamma receptor 2 (interferon gamma transducer 1)
heterogeneous nuclear ribonucleoprotein K
splicing factor, arginine/serine-rich 1
Rho GDP dissociation inhibitor (GDI) alpha
follistatin-like 1
golgi phosphoprotein 3 (coat-protein)
sphingomyelin synthase 1
glyceraldehyde-3-phosphate dehydrogenase
HNRNPA3
SYPL1
MLANA
DNAJA3
COL5A2
UBE2G1
CHMP4C
HPGD
OBFC2A
RALGAPA1
ATP1A2
GALNT1
heterogeneous nuclear ribonucleoprotein A3
synaptophysin-like 1
DnaJ (Hsp40) homolog, subfamily A, member 3
collagen, type V, alpha 2
ubiquitin-conjugating enzyme E2G 1 (UBC7 homolog, yeast)
chromatin modifying protein 4C
hydroxyprostaglandin dehydrogenase 15-(NAD)
oligonucleotide/oligosaccharide-binding fold containing 2A
Ral GTPase activating protein, alpha subunit 1 (catalytic)
ATPase, Na+/K+ transporting, alpha 2 polypeptide
UDP-N-acetyl-alpha-D-galactosamine: N-acetylgalactosaminyltransferase 1
-1,354
-1,353
-1,352
-1,35
-1,346
-1,336
-1,329
-1,329
-1,324
-1,323
-1,318
-1,318
-1,316
-1,313
-1,311
-1,304
-1,3
-1,3
-1,297
-1,297
-1,295
-1,295
-1,295
-1,293
-1,289
-1,286
-1,284
-1,284
-1,283
-1,282
-1,279
-1,276
-1,274
-1,274
-1,274
-1,271
-1,268
-1,268
-1,265
-1,261
-1,259
-1,258
-1,258
-1,256
-1,254
-1,254
-1,253
-1,253
-1,252
-1,252
-1,251
-1,251
-1,25
-1,249
-1,247
-1,245
-1,244
-1,242
-1,241
-1,241
-1,24
-1,239
-1,237
-1,237
-1,237
-1,237
Cfl2
Rock1
Arl6ip5
Prrx1
Cpsf2
Gata6
Fgf1
Sorbs1
Rab10
Pik3ca
Hccs
Cul3
Aspn
Rnf114
Ythdf3
Ttll5
Sepx1
Hif1a
Tmem65
Gyk
Eif4a1
Mrps22
Col4a5
Tmod1
Clip1
Stag2
Cuzd1
Nolc1
Gucy1b3
Eif2s3y
Cab39
Dnajb6
Lactb2
Nckap1
Twsg1
Ahnak
Smpx
Acadl
Ube2d3
Synpo2
Gnai3
Tiprl
Txnl1
Ccar1
Gt(ROSA)2
6SorEaf1
Arih1
Gatm
Idh1
Sar1b
Eif3h
Cand2
Pabpc1
Tmem144
Hars
Adsl
Mrpl9
Yipf5
Anp32b
Fbxo3
Arl1
Tcea1
Oxct1
Vim
Crk
Pde3a
CFL2
ROCK1
ARL6IP5
PRRX1
CPSF2
GATA6
FGF1
SORBS1
RAB10
PIK3CA
HCCS
CUL3
ASPN
RNF114
YTHDF3
TTLL5
SEPX1
HIF1A
TMEM65
cofilin 2 (muscle)
Rho-associated, coiled-coil containing protein kinase 1
ADP-ribosylation-like factor 6 interacting protein 5
paired related homeobox 1
cleavage and polyadenylation specific factor 2, 100kDa
GATA binding protein 6
fibroblast growth factor 1 (acidic)
sorbin and SH3 domain containing 1
RAB10, member RAS oncogene family
phosphoinositide-3-kinase, catalytic, alpha polypeptide
holocytochrome c synthase
cullin 3
asporin
ring finger protein 114
YTH domain family, member 3
tubulin tyrosine ligase-like family, member 5
selenoprotein X, 1
hypoxia inducible factor 1, alpha subunit
transmembrane protein 65
EIF4A1
MRPS22
COL4A5
TMOD1
CLIP1
STAG2
CUZD1
NOLC1
GUCY1B3
eukaryotic translation initiation factor 4A1
mitochondrial ribosomal protein S22
collagen, type IV, alpha 5
tropomodulin 1
CAP-GLY domain containing linker protein 1
stromal antigen 2
CUB and zona pellucida-like domains 1
nucleolar and coiled-body phosphoprotein 1
guanylate cyclase 1, soluble, beta 3
CAB39
DNAJB6
LACTB2
NCKAP1
TWSG1
AHNAK
SMPX
ACADL
UBE2D3 ‡
SYNPO2
GNAI3
TIPRL
TXNL1
CCAR1
calcium binding protein 39
DnaJ (Hsp40) homolog, subfamily B, member 6
lactamase, beta 2
NCK-associated protein 1
twisted gastrulation homolog 1 (Drosophila)
AHNAK nucleoprotein
small muscle protein, X-linked
acyl-CoA dehydrogenase, long chain
ubiquitin-conjugating enzyme E2D 3 (UBC4/5 homolog, yeast)
synaptopodin 2
guanine nucleotide binding protein, alpha inhibit.activity polypeptide 3 TIP41, TOR
signaling pathway regulator-like (S. cerevisiae)
thioredoxin-like 1
cell division cycle and apoptosis regulator 1
EAF1
ARIH1
GATM
IDH1
SAR1B
EIF3H
CAND2
PABPC1
TMEM144
HARS
ADSL
MRPL9
YIPF5
ANP32B
FBXO3
ARL1
TCEA1
OXCT1
VIM
CRK
PDE3A
ELL associated factor 1
ariadne homolog, ubiquitin-conjug.enz.e E2 binding protein, 1 Droso)
glycine amidinotransferase (L-arginine:glycine amidinotransferase)
isocitrate dehydrogenase 1 (NADP+), soluble
SAR1 homolog B (S. cerevisiae)
eukaryotic translation initiation factor 3, subunit H
cullin-associated and neddylation-dissociated 2 (putative)
poly(A) binding protein, cytoplasmic 1
transmembrane protein 144
histidyl-tRNA synthetase
adenylosuccinate lyase
mitochondrial ribosomal protein L9
Yip1 domain family, member 5
acidic (leucine-rich) nuclear phosphoprotein 32 family, member B
F-box protein 3
ADP-ribosylation factor-like 1
transcription elongation factor A (SII), 1
3-oxoacid CoA transferase 1
vimentin
v-crk sarcoma virus CT10 oncogene homolog (avian)
phosphodiesterase 3A, cGMP-inhibited
-1,235
-1,235
-1,234
-1,232
-1,231
-1,231
-1,227
-1,226
-1,224
-1,223
-1,221
-1,221
-1,21
-1,21
-1,209
-1,209
-1,208
-1,207
-1,203
-1,202
-1,201
-1,2
-1,2
-1,198
-1,192
-1,188
-1,187
-1,187
-1,187
-1,186
-1,175
-1,174
-1,171
-1,167
-1,162
-1,16
-1,153
1,144
1,161
1,165
1,173
1,174
1,176
1,18
1,187
1,192
1,198
1,204
1,206
Vapb
Ubr3
Rpsa
Crtap
Zdhhc3
Cxadr
Ptpla
Pdha1
Pdgfa
Plagl1
Mtmr6
Rnase4
Ptges3
Dda1
Nsmce2
Fam63a
Slc25a11
Smyd2
Ctnnb1
Col9a3
Baz1b
Rraga
Lass2
Cryab
Uba5
Hnrnpd
Degs1
Qpct
Cd68
Ppp2r1a
Fbxo8
Ltbp1
Cbr2
App
Ankrd46
Oxsr1
Gtl3
Stk39
Asnsd1
Hrc
Sorbs3
P4ha1
Syngr1
Ndufs2
Cnn2
Ubxn1
B3gat3
Asah2
Gzma
VAPB
UBR3
RPSA
CRTAP
ZDHHC3
CXADR
PTPLA
PDHA1 §
PDGFA
PLAGL1
MTMR6
RNASE4
PTGES3 ||
DDA1
NSMCE2
FAM63A
SLC25A11
SMYD2
CTNNB1
COL9A3
BAZ1B
RRAGA
LASS2
CRYAB
UBA5
HNRNPD
DEGS1
QPCT #
CD68
PPP2R1A
FBXO8
LTBP1
VAMP (vesicle-associated membrane protein)-associated protein B,C
ubiquitin protein ligase E3 component n-recognin 3 (putative)
ribosomal protein SA
cartilage associated protein
zinc finger, DHHC-type containing 3
coxsackie virus and adenovirus receptor
protein tyrosine phosphatase-like(proline instead of catalytic arginine),A
pyruvate dehydrogenase (lipoamide) alpha 1
platelet-derived growth factor alpha polypeptide
pleiomorphic adenoma gene-like 1
myotubularin related protein 6
ribonuclease, RNase A family, 4
prostaglandin E synthase 3 (cytosolic)
DET1 and DDB1 associated 1
non-SMC element 2, MMS21 homolog (S. cerevisiae)
family with sequence similarity 63, member A
solute carrier family 25 (mitochondrial oxoglutarate carrier), member 11
SET and MYND domain containing 2
catenin (cadherin-associated protein), beta 1, 88kDa
collagen, type IX, alpha 3
bromodomain adjacent to zinc finger domain, 1B
Ras-related GTP binding A
LAG1 homolog, ceramide synthase 2
crystallin, alpha B
ubiquitin-like modifier activating enzyme 5
heterogeneous nuclear ribonucleoprotein D
degenerative spermatocyte homolog 1, lipid desaturase (Drosophila)
glutaminyl-peptide cyclotransferase
CD68 molecule
protein phosphatase 2, regulatory subunit A, alpha
F-box protein 8
latent transforming growth factor beta binding protein 1
APP
ANKRD46
OXSR1
amyloid beta (A4) precursor protein
ankyrin repeat domain 46
oxidative-stress responsive 1
1,207
1,21
1,216
1,22
1,222
1,222
1,229
1,232
1,234
1,238
1,248
1,25
1,254
1,259
1,266
1,267
Dcaf11
Spr
Mif
Trappc6a
Dctn4
Egln3
Txn1
Uxt
Lgr5
Cript
Asb11
Adrbk1
Cd47
Gys1
Pitpnc1
Cecr6
DCAF11
SPR
MIF
TRAPPC6A
DCTN4
EGLN3
STK39
ASNSD1
HRC
SORBS3
P4HA1
SYNGR1
NDUFS2
CNN2
UBXN1
B3GAT3
ASAH2
GZMA
UXT
LGR5
CRIPT
ASB11
ADRBK1
CD47
GYS1
PITPNC1
CECR6
serine threonine kinase 39 (STE20/SPS1 homolog, yeast)
asparagine synthetase domain containing 1
histidine rich calcium binding protein
sorbin and SH3 domain containing 3
prolyl 4-hydroxylase, alpha polypeptide I
synaptogyrin 1
NADHdehydrogenase Fe-S protein 2,(NADH-co Q reductase)
calponin 2
UBX domain protein 1
beta-1,3-glucuronyltransferase 3 (glucuronosyltransferase I)
N-acylsphingosine amidohydrolase (non-lysosomal ceramidase) 2
granzyme A (granzyme 1, cytotoxic T-lymphocyte-associated serine esterase 3)
DDB1 and CUL4 associated factor 11
sepiapterin reductase (7,8-dihydrobiopterin:NADP+ oxidoreductase)
macrophage migration inhibitory factor (glycosylation-inhibiting factor)
trafficking protein particle complex 6A
dynactin 4 (p62)
egl nine homolog 3 (C. elegans)
ubiquitously-expressed transcript
leucine-rich repeat-containing G protein-coupled receptor 5
cysteine-rich PDZ-binding protein
ankyrin repeat and SOCS box-containing 11
adrenergic, beta, receptor kinase 1
CD47 molecule
glycogen synthase 1 (muscle)
phosphatidylinositol transfer protein, cytoplasmic 1
cat eye syndrome chromosome region, candidate 6
1,269
1,271
1,285
1,285
1,299
H2-Ke6
Txnrd2
Pdp2
Bag1
Ctdsp1
TXNRD2
PDP2
BAG1
CTDSP1
1,302
1,32
1,34
1,394
1,452
1,492
1,766
Gatad2a
Dhrs3
Ppara
Grm1
Dpf2
Srrm1
Slc41a3
GATAD2A
DHRS3
PPARA
GRM1
DPF2
SRRM1
SLC41A3 **
thioredoxin reductase 2
pyruvate dehyrogenase phosphatase catalytic subunit 2
BCL2-associated athanogene
CTD (carboxy-terminal domain, RNA polymerase II, polypeptide A) small
phosphatase 1
GATA zinc finger domain containing 2A
dehydrogenase/reductase (SDR family) member 3
peroxisome proliferator-activated receptor alpha
glutamate receptor, metabotropic 1
D4, zinc and double PHD fingers family 2
serine/arginine repetitive matrix 1
solute carrier family 41, member 3