SUPPORTING INFORMATION
Figure S1.
Light microscopy of transverse and longitudinal semi-thin sections of gastrocnemius
muscles from young (a, c) and aged (b, d) rats. Signs of aging processes are represented by a
splitting fiber (b, arrowheads point the invaginated sarcolemma) and an area of myofibrillar
degeneration (d, arrow). Toluidine blue staining. Scale bar = 20 µm.
Figure S2.
Light microscopy of transverse and longitudinal semi-thin sections of triceps muscles from
young (a, c) and aged (b, d) rats. Signs of aging processes are represented by internal migration
of nuclei (b, arrows), areas of disarray of myofibrils (d, upper fiber) and myofibrillar degeneration (d,
arrowheads). Toluidine blue staining. Scale bar = 20 µm.
Figure S3.
a-d) Electron microscopy images of muscles
from aged rats. In the gastrocnemius muscle
(a) are evident lamellar formations, parallel
arranged and close to the nucleus (at high
magnification in the inset). In the triceps muscle
(b) are evident small foci of myofibrillar loss (red
asterisks). Scale bar = 1 µm; inset scale bar =
0.25 µm. In the gastrocnemius (c) is present a
lipofuscin granule (arrow); in the triceps (d) are
present lipid droplets (arrowheads). Scale bar =
1 µm.
e-h) Electron microscopy images of
subsarcolemmal
and
intermyofibrillar
mitochondria. Gastrocnemius (e,f,g) and
triceps (h) muscles from aged rats show some
age related modifications of mitochondria. In
aged gastrocnemius (e), some subsarcolemmal
mitochondria, appear larger (arrows) than
others. An autophagosome is marked by
asterisk. (f) an extended intermyofibrillar
network
of
elongated
mitochondria
(arrowheads) is evident in this muscle. (g)
Evidence of a mitochondrion with crystalloid
inclusions,
suggestive
of
morphological
changes, between normal myofibrils (higher
magnification of a small detail is shown in the
inset). (h) Subsarcolemmal accumulation of
mitochondria and a lipid droplet (arrowhead) in
aged triceps muscle. Scale bar = 1 µm.
Figure S4.
SDS-PAGE of young (Y) and aged (A) gastrocnemius (Gas) and triceps (Tri) muscle extracts.
Samples (50 µg) were loaded on a 6-14% gradient polyacrylamide gel and stained using SYPRO
Orange protein gel stain (Molecular probes). Low and High Molecular weight markers (Ge
Healthcare) were run in separate lanes.
Figure S5.
Gastrocnemius (A) and triceps (B) muscles protein profiling by 2D-DIGE. Protein extracts (50
µg) from each set type were labeled with 400 pmol Cy5 dye (CyDye; GE healthcare), while an
internal standard, generated by pooling together an aliquot of all the samples, was labeled with the
Cy3 dye. Individual samples (40 µg) were combined with an equal amount of internal standard and
separated on 24 cm, nonlinear pH 3–10 gradient IPGstrips (first dimension) and on 20 × 25 cm2,
12% T, 2.5% C polyacrylamide gels (second dimension). Protein spots significantly changed
(Student’s t-test, p < 0.01) in 22-month-old gastrocnemius and triceps extracts compared with their
3-month-old controls and identified by MALDI-ToF mass spectrometry are indicated in
representative 2D maps and numbered accordingly to Supplementary Table S2A and S2B listing.
Figure S6.
Validation of MALDI-ToF MS identified proteins by immunoblotting.
Immunoblotting of selected proteins (identified by proteomic analysis) in gastrocnemius and triceps
muscles from aged rats versus their respective young controls. Statistical analysis was performed
by Student’s T-Test (n=6; * p<0.05). The data were normalized against the total amount of loaded
proteins stained with Sypro Ruby. GAPDH: glyceraldehyde 3-phosphate dehydrogenase; LDHA: Llactate dehydrogenase A chain; CKM: creatine kinase M-type; ACO2: aconitase 2; ATP5B: ATP
synthase subunit beta; CRYAB: alpha-crystallin B chain; CAR3: carbonic anhydrase 3; VDAC1:
voltage-dependent ion channel 1.
Figure S7.
Biochemical analysis of myogenic regulatory transcription factors (MRFs). Upper panel:
Representative immunoblot images of Pax7, MyoD and myogenin in gastrocnemius (Gas) and
triceps (Tri) muscles of aged rats (A) and young controls (Y). Lower panel: Histograms of protein
expression (normalized to their respective controls) in gastrocnemius and triceps muscles of aged
rats (mean ± SD). *Significant differences (Student’s t-test, n = 3, p < 0.05) of aged vs. control rats.
Table S1. MIAPE Standards Data
MIAPE-GE Gel Electrophoresis Supplementary Information
General Features
•
22-month-old (aged) and 3-month-old (young) Sprague-Dawley male rats (Charles River,
Calco, Italy) were used in this study. All the procedures involving animals and their care
were conducted in conformity with the institutional procedures in compliance with national
(D.L. No. 116, G.U. Suppl. 40, Feb. 18, 1992, Circolare No. 8, G.U., 14 Luglio 1994) and
international regulations (EEC Council Directive 86/609, OJ L 358, 1 DEC.12, 1987; NIH
Guide for the Care and use of Laboratory Animals, U.S. National Research Council,
1996).
•
Experiments conducted at the Università degli Studi di Milano, Dipartimento di Scienze
Biomediche per la Salute, Segrate (MI), Italia.
•
Electrophoresis type: 2-D DIGE.
•
Images: available upon request.
•
Image analysis software: DeCyder version 6.5 (GE Healthcare).
•
Statistical analysis software: EDA module version 1.0 (GE Healthcare).
Gel analysis design
•
Animals: 12 male rats (6 young and 6 aged).
•
Samples: gastrocnemius and triceps muscles from left and right fore- and hind-limbs.
•
Sample preparation: an aliquot of each frozen muscle was suspended in lysis buffer (urea
7M, thiourea 2M, CHAPS 4%, Tris 30mM, and PMSF 1mM) and solubilized by sonication
on ice. Proteins were selectively precipitated using PlusOne 2D-Clean-up kit (GE
Healthcare) in order to remove nonprotein impurities and re-suspended in lysis buffer.
Protein extract was adjusted to pH 8.5 by the addition of NaOH 1M solution, and sample
concentrations were determined using PlusOne 2-D Quant kit (GE Healthcare).
•
Experimental groups: 4 (aged gastrocnemius, aged triceps, young gastrocnemius, young
triceps) – 12 samples for each experimental group. 6 aged and 6 young samples for each
muscle type were used for DIGE analysis. Each sample was Cy5-labelled and run in
duplicate.
•
Internal standard: Cy3-labelled pool of equal protein amounts from all samples run on
each analytic gel.
Image pre-processing
•
Type: Images were scanned on the Typhoon 9200 Imager (GE Healthcare).
•
Software: ImageQuant TL (GE Healthcare).
Settings:
•
•
•
Analytic (quantitative) gels: laser voltages were optimized for each fluorophore prior to
scanning to avoid signal saturation. Identical laser settings were then used to scan each
gel.
Preparative (picking/protein identification) gel: fixed with ethanol/acetic acid and poststained with Deep Purple, scanned independently from analytic gels.
All gels were scanned at 100μm resolution.
Data extraction process
•
Input images: available upon request.
•
Feature detection: DeCyder version 6.5, DIA module (GE Healthcare)
•
Automatic detection with estimation of 5000 protein spots
•
Exclusion filter applied to spots with the following features
•
Slope > 1.2
•
Area < 200
•
Volume < 10000
•
•
•
•
•
•
•
•
DIA workspaces were manually edited to eliminate gel artefacts and to reinclude any
incorrectly excluded spots.
Matching algorithm: DeCyder version 6.5, BVA module (GE Healthcare)
Automatic matching with 50 manually identified landmark protein spots per gel
Spot matches were manually edited
Match on 90% of gels required.
Feature quantitation: Decyder version 6.5
Background subtraction: DeCyder version 6.5
Normalization: DeCyder version 6.5.
Data analysis
•
PCA: DeCyder EDA module version 1.0.
•
Statistical analysis: DeCyder EDA module version 1.0.
•
Type: Student’s T-test (p<0.05); input data: log standardized abundance.
•
Power analysis: Lenth power tool; input data: biological variance calculated from
standardized spots volumes.
MIAPE MS Supplementary Information
General features
•
Instrument used: MALDI ULTRAFLEX III (Bruker Daltonics).
•
Software used: FlexAnalysis v. 3.0 (Bruker Daltonics); MASCOT v 2.2 database
Input data and parameters
•
Type of MS data: MS spectra.
•
Databases queried: MASCOT v2.2, NCBInr 20090430 database (8483808 sequences;
2914572939 residues).
•
Taxonomical restrictions: Rodentia.
•
Cleavage agent: Trypsin.
•
Allowed number of missed cleavages: 1.
•
Permissible amino acid modifications: oxidation (variable) and carbamidomethylation
(fixed).
•
Mass exclusion tolerance: 30 ppm.
The output from the procedure
•
Data for identified proteins: See Supporting Information Table S2A, S2B and annotated
spectra of identified proteins.
Protein identification methods
Spots of interest were excised from gel using the Ettan spot picker robotic system (GE
Healthcare), destained in 50% methanol/50mM ammonium bicarbonate (AMBIC), reduced,
alkylated and incubated with 30 µL of 6 ng/µL trypsin (Promega) dissolved in 10 mM AMBIC
for 16 h at 37°C. Released peptides were subjected to reversed-phase chromatography (ZipTip C18 micro, Millipore), eluted with 50%ACN/1% formic acid. Peptides mixture (1 µL) was
diluted in an equal volume of 10 mg/mL CHCA matrix dissolved in 70% ACN/30% citric acid
and processed on a Ultraflex III MALDI-ToF/ToF (Bruker Daltonics) mass spectrometer. MS
was performed at an accelerating voltage of 25 kV and spectra were externally calibrated
using Peptide Mix calibration mixture (Bruker Daltonics); 1000 laser shots were taken per
spectrum. Spectra were processed by the FlexAnalysis software v. 3.0 (Bruker Daltonics)
setting the signal-to-noise threshold value to 6 and search was carried out by correlation of
uninterpreted spectra to Rodentia entries in NCBInr 20090430 database (8.483.808
sequences; 2.914.572.939 residues). Proteins were identified by comparing the digest peaks
with a computer-generated database of tryptic peptides from known proteins using MASCOT,
which utilizes a robust probabilistic scoring algorithm. With regard to MASCOT parameters,
the significance threshold was set at p-value<0.05. One missed cleavage per peptide was
allowed and carbamidomethylation was set as a fixed modification while methionine oxidation
as a variable modification. Mass tolerance was set at 30 ppm.
Table S2.
Protein Identification by MALDI-ToF MS in gastrocnemius (A) and triceps (B) muscles. For
each identified spot, characterized by a unique spot number assigned during the matching process
by the DeCyder software, mass spectrometry parameters (theoretical MW and pI, MASCOT score,
protein coverage, n° matched over searched peaks) and statistical analysis parameters (t-test pvalue, average ratio aged/young, % variation) are reported, along with recommended protein name,
gene symbol, and UniProt Knowledgebase accession number.
Table S2A. List of changed proteins in aged vs. young gastrocnemius muscle, identified by MALDI-ToF MS.
2D map reference
Protein
Gene
name
UniProtKB entry
Theoretical MW
Theorecical pI
Mascot
Coverage
Matched/Searched
Peptides
T-test p-value
Av. Ratio
% variation
Anaerobic metabolism:
1
Aldehyde reductase 1
Akr1b1
P07943
35,8
6,3
112
32,9
9/21
1,90E-05
-1,43
-30
2
Alpha enolase
Eno1
P04764
47,1
6,2
93
23
10/20
0,00015
-1,51
-34
3
Beta enolase
Eno3
P15429
47
6,9
93
21
7/16
4,60E-05
-1,37
-27
4
Beta enolase
Eno3
P15429
47
7,8
148
39,2
13/21
4,60E-05
-1,39
-28
5
Beta enolase
Eno3
P15429
47
7,8
138
32,9
12/32
2,10E-05
-1,35
-26
6
Beta enolase
Eno3
P15429
47
7,8
188
38,2
16/27
3,60E-05
-1,37
-27
7
Fructose-bisphosphate aldolase A
Aldoa
P05065
39,3
9,2
202
38,5
16/34
0,00083
-1,29
-22
8
Fructose-bisphosphate aldolase A
Aldoa
P05065
39,3
9,2
248
55,2
18/32
0,0049
-1,27
-21
9
Glyceraldehyde 3-phosphate dehydrogenase
Gapdh
P04797
35,8
9
157
35,1
10/15
2,60E-06
-1,82
-45
10
Glyceraldehyde 3-phosphate dehydrogenase
Gapdh
P04797
35,8
9
100
29,7
8/22
2,40E-07
-1,83
-45
11
Glyceraldehyde 3-phosphate dehydrogenase
Gapdh
P04797
35,8
9
113
39,6
11/28
2,40E-05
-1,89
-47
12
Glyceraldehyde 3-phosphate dehydrogenase
Gapdh
P04797
35,8
9
95
30,6
9/26
0,00036
-1,37
-27
13
Glycerol-3-phosphate dehydrogenase [NAD+], cytoplasmic
Gpd1
O35077
37,4
6,2
119
29,8
11/19
0,0028
-1,17
-15
14
Glycogen phosphorylase, muscle form
Pygm
P09812
97,2
6,7
200
25,9
19/26
0,0002
-1,5
-33
15
Glycogen phosphorylase, muscle form
Pygm
P09812
97,2
6,7
151
20,5
18/30
6,30E-05
-1,53
-35
16
Glycogen phosphorylase, muscle form
Pygm
P09812
97,2
6,7
304
38,4
26/32
0,0001
-1,57
-36
17
L-lactate dehydrogenase A chain
Ldha
P04642
36,4
9,3
181
41
18/26
8,70E-05
-1,53
-35
18
L-lactate dehydrogenase A chain
Ldha
P04642
36,4
9,3
178
36,4
15/22
5,50E-05
-1,57
-36
19
L-lactate dehydrogenase A chain
Ldha
P04642
36,4
9,3
180
37
14/19
0,0019
-1,58
-37
20
Phosphoglucomutase
Pgm1
P38652
61,4
6,1
113
26,3
12/27
5,50E-05
-1,41
-29
21
Phosphoglycerate mutase 2
Pgam2
P16290
28,7
9,5
157
48,2
12/22
0,0064
-1,29
-22
22
Phosphoglycerate mutase 2
Pgam2
P16290
28,7
9,5
151
44,3
11/20
3,40E-05
-1,38
-28
23
Phosphoglycerate mutase 2
Pgam2
P16290
28,7
9,5
153
44,3
12/22
5,00E-05
-1,39
-28
24
Pyruvate Kinase, isozymes M1/M2
Pkm
P11980
57,8
6,7
130
28,2
12/23
0,0006
-1,39
-28
25
Triosephosphate isomerase
Tpi1
P48500
26,8
7,7
161
51
11/22
0,00011
-1,14
-12
26
Creatine kinase, M chain
Ckm
P00564
43
6,6
223
38,3
18/22
0,0026
-1,27
-21
27
Creatine kinase, M chain
Ckm
P00564
43
6,6
100
31,2
9/24
0,00079
-1,32
-24
28
Creatine kinase, sarcomeric mitochondrial
Ckmt2
P09605
47,4
9,5
199
42,5
19/30
5,80E-05
-1,19
-16
29
Adenylate kinase isoenzyme 1
Ak1
P39069
21,6
8,8
140
52,6
12/23
0,0011
-1,29
-22
30
Nucleoside diphosphate kinase B
Nme2
P19804
17,3
7,8
186
84,2
11/18
0,0031
-1,16
-14
Energy balance:
TCA cycle:
31
Aconitate hydratase, mitochondrial,
Aco2
Q9ER34
85,4
8,7
117
22,6
14/36
0,00011
1,49
49
32
Aconitate hydratase, mitochondrial,
Aco2
Q9ER34
85,4
8,7
373
51,9
33/58
5,00E-06
1,34
34
33
Aconitate hydratase, mitochondrial,
Aco2
Q9ER34
85,4
8,7
134
23,3
17/45
0,001
1,24
24
34
Aconitate hydratase, mitochondrial,
Aco2
Q9ER34
85,4
8,7
252
34,7
25/43
0,00025
1,29
29
35
Dihydrolipoamide dehydrogenase, mitochondrial
Dld
Q6P6R2
54
9
154
26,9
14/33
0,0001
1,23
23
36
Dihydrolipoyllysine-residue acetyltransferase component of pyruvate dehydrogenase complex
Dlat
P08461
53,2
9,3
85
16,6
9/15
0,001
1,21
21
37
Isocitrate dehydrogenase [NAD] subunit alpha, mitochondrial precursor
Idh3a
Q99NA5
39,6
6,5
155
34,4
17/41
9,10E-06
1,29
29
38
Isocitrate dehydrogenase [NAD] subunit alpha, mitochondrial precursor
Idh3a
Q99NA5
39,6
6,5
81
23
9/26
0,00066
1,17
17
39
Isocitrate dehydrogenase [NADP], mitochondrial [precursor]
Idh2
P56574
50,9
9,6
214
45,4
22/49
0,0003
1,5
50
40
Pyruvate dehydrogenase E1 component beta subunit, mitochondrial
Pdhb
P49432
39
6,2
130
41,5
13/26
0,00056
1,34
34
41
Succinyl-CoA ligase [ADP-forming] beta-chain, mitochondrial, [precursor],
Sucla2
F1LM47
46,2
5,6
92
22,3
11/27
0,0021
1,19
19
42
Succinyl-CoA: 3-ketoacid-coenzyme A transferserase 1, mitochondrial [precursor]
Oxct1
B2GV06
56
9,5
116
29
11/21
0,00018
1,29
29
43
ATP synthase alpha chain, mitochondrial
Atp5a1
P15999
59,7
9,7
92
22,2
11/26
5,60E-06
1,34
34
44
ATP synthase alpha chain, mitochondrial
Atp5a1
P15999
59,7
9,7
266
42,2
20/24
1,20E-05
1,27
27
45
ATP synthase beta chain, mitochondrial
Atp5b
P10719
56,1
5,1
161
36,1
15/29
9,70E-05
1,32
32
46
ATP synthase beta chain, mitochondrial
Atp5b
P10719
56,1
5,1
261
53,9
27/40
1,40E-05
1,36
36
47
ATP synthase D chain, mitochondrial
Atp5h
P31399
18,8
5,7
103
59,6
9/39
0,0021
1,41
41
48
Ubiquinol-cytochrome C reductase complex co. protein I, mitochondrial
Uqcrc1
Q9CZ13
52,8
5,5
170
24,4
11/16
0,00031
1,52
52
49
Actin, alpha skeletal muscle
Acta1
P68136
37,8
5,4
145
36,3
13/24
0,00025
1,26
26
50
Cofilin, muscle isoform
Cfl2
M0RC65
18,7
8,8
76
31,3
5/8
0,0053
1,64
64
51
Desmin
Des
P48675
53,4
5,1
325
49
28/39
5,50E-05
1,42
42
52
Desmin
Des
P48675
53,4
5,1
309
52
30/46
0,0049
1,28
28
53
Desmin
Des
P48675
53,4
5,1
307
58
29/57
0,0021
1,29
29
54
Myosin binding protein C, fast-type
Mybpc2
D3ZA38
127,2
6
135
17,2
16/25
1,10E-05
-1,28
-22
55
Myosin binding protein C, fast-type
Mybpc2
D3ZA38
127,2
6
126
17,4
16/30
1,40E-05
-1,31
-24
56
Myosin binding protein C, slow type
Mybpc1
Q63518
126,1
5,6
251
22,8
29/36
0,00011
1,64
64
57
Myosin binding protein C, slow type
Mybpc1
Q63518
126,1
5,6
141
16,2
17/25
7,50E-05
1,7
70
58
Myosin heavy chain IIB [Fragment]
Myh4
Q9JHR4
61
5,3
286
44,6
26/37
0,0027
-1,43
-30
59
Myosin heavy chain IIB [Fragment]
Myh4
Q9JHR4
61
5,3
201
32,5
16/19
0,0002
-1,68
-40
60
Myosin light chain 1, skeletal muscle isoform
Myl1
P02600
20,7
4,8
183
63,5
14/25
0,00045
-1,45
-31
61
Myosin-binding protein H
Mybph
O88599
52,6
6
113
33,5
12/38
0,00084
-1,53
-35
62
Myosin-binding protein H
Mybph
O88599
52,6
6
73
18,4
7/19
3,50E-05
-1,57
-36
63
Myozenin-1
Myoz1
D4A7U8
31,4
9,1
110
34,5
9/12
0,00029
-1,23
-19
64
PDZ-LIM protein cypher 2s
Ldb3
Q5XIG1
35,1
9,6
187
47,4
16/30
0,00041
-1,15
-13
65
PDZ-LIM protein cypher 2s
Ldb3
Q5XIG1
35,1
9,6
143
35,5
13/27
0,0066
-1,19
-16
Respiratory chain:
Structural proteins:
66
Skeletal muscle-specific alpha-actinin 3
Actn3
Q8R4I6
102,9
5,2
361
41,1
37/49
67
Tropomyosin 1 alpha chain
68
Tubulin beta-2C chain
69
70
0,001
-1,09
-8
Tpm1
P04692
32,7
4,6
Tubb4b
Q6P9T8
49,8
4,6
189
44
246
33,7
17/35
0,0042
-1,3
-23
23/26
6,80E-05
1,9
90
Myoglobin
Mb
Q9QZ76
17,1
Myoglobin
Mb
Q9QZ76
17,1
8,9
127
8,9
159
55,2
8/11
0,0031
1,43
43
63
11/20
0,0039
1,35
35
71
Serotransferrin
Tf
P12346
72
Serotransferrin
Tf
P12346
76,3
7,2
76,3
7,2
266
28,9
19/21
0,00018
1,44
44
182
30,2
17/28
0,00012
1,33
73
Serum albumin
Alb
P02770
68,7
33
6,1
180
33,9
18/37
3,00E-06
1,61
74
Serum albumin
Alb
P02770
61
68,7
6,1
334
54,3
29/49
5,70E-06
1,77
77
0,0024
1,16
16
Transport proteins:
Cell Signaling and Stress Proteins:
75
14-3-3 protein gamma-subtype
Ywhag
P61983
28,3
4,7
110
27,5
10/14
76
Alpha crystallin B chain
Cryab
P23928
19,9
6,9
169
62,6
12/26
0,00018
3,17
217
77
Carbonic anhydrase III
Car3
P14141
29,4
7,1
197
54,2
14/25
1,50E-06
7,02
602
78
Stress-70 protein, mitochondrial precursor
Hspa9
P48721
73,8
5,9
205
43,2
24/47
0,00042
1,94
94
79
Mitochondrial inner membrane protein
Immt
Q3KR86
67,1
5,5
322
43,3
29/45
0,0025
1,21
21
80
Mitochondrial inner membrane protein
Immt
Q3KR86
67,1
5,5
154
17,4
11/14
0,00012
1,93
93
81
Ppp2r1a protein
Ppp2r1a
Q5XI34
65,3
4,9
107
19,2
11/18
0,0055
1,22
22
82
Prohibitin [Mus musculus]
Phb
P67779
29,8
5,5
178
58,1
12/22
0,004
4,7
370
83
Voltage-dependent anion-selective channel protein 1
Vdac1
Q9Z2L0
30,7
8,6
185
52,7
14/33
0,00017
1,39
39
84
Voltage-dependent anion-selective channel protein 1
Vdac1
Q9Z2L0
30,7
8,6
106
38,2
9/31
0,00027
1,26
26
85
Voltage-dependent anion-selective channel protein 2
Vdac2
P81155
31,7
8,7
96
29,2
6/11
1,00E-05
1,81
81
86
Voltage-dependent anion-selective channel protein 2
Vdac2
P81155
31,7
8,7
91
29,2
7/18
3,80E-05
1,41
41
Table S2B. List of changed proteins in aged vs. young triceps muscle, identified by MALDI-ToF MS.
2D map reference
Protein
Gene
name
UniProtKB entry
Theoretical
MW
Theorecical pI
Mascot
Coverage
Matched/Searched
Peptides
T-test
Av. Ratio
% variation
Anaerobic metabolism:
1
Aldehyde reductase 1
Akr1b1
P07943
35,7
6,9
91
34,5
8/28
1,85E-03
-1,22
-18
2
Alpha enolase
Eno1
P04764
47,1
6,2
142
40,3
15/30
3,10E-04
-3,41
-71
3
Beta enolase
Eno3
P15429
47
6,9
87
23,7
9/24
4,80E-05
-1,26
-21
4
Beta enolase
Eno3
P15429
47
6,9
217
50,9
19/27
1,81E-05
-1,36
-26
5
Beta enolase
Eno3
P15429
47
6,9
220
53,3
19/30
9,52E-05
-1,25
-20
6
Fructose-bisphosphate aldolase A
Aldoa
P05065
39,3
9,2
221
52,5
18/31
7,62E-05
-1,22
-18
7
Fructose-bisphosphate aldolase A
Aldoa
P05065
39,3
9,2
132
49,2
13/33
1,21E-04
-1,32
-24
8
Glyceraldehyde 3-phosphate dehydrogenase
Gapdh
P04797
35,8
9
126
34,5
10/24
8,76E-04
-1,33
-25
9
Glyceraldehyde 3-phosphate dehydrogenase
Gapdh
P04797
35,8
9
113
39
13/38
2,64E-04
-1,22
-18
10
Glycerol-3-phosphate dehydrogenase [NAD+], cytoplasmic
Gpd1
O35077
37,4
6,2
192
48,1
17/24
7,99E-05
-1,34
-25
11
Phosphoglycerate mutase 2
Pgam2
P16290
28,7
9,5
166
52,2
13/35
1,20E-04
1,35
35
12
Phosphoglycerate mutase 2
Pgam2
P16290
28,7
9,5
117
39,9
10/24
3,58E-03
1,25
25
13
Pyruvate Kinase, isozymes M1/M2
Pkm
P11980
57,8
6,7
249
44,4
22/32
1,66E-04
-1,27
-21
14
Triosephosphate isomerase
Tpi1
P48500
26,8
7,7
160
51
11/25
2,83E-05
1,41
41
15
Creatine kinase, M chain
Ckm
P00564
43
6,6
238
53,8
19/36
9,62E-05
-1,28
-22
16
Creatine kinase, M chain
Ckm
P00564
43
6,6
217
42
15/20
2,30E-05
-1,3
-23
17
Adenylate kinase isoenzyme 1
Ak1
P39069
21,6
8,8
120
47,9
9/16
0,0000112
-1,21
-17
18
Dihydrolipoyllysine-residue acetyltransferase component of pyruvate dehydrogenase complex
Dlat
P08461
49
9,9
135
31,9
14/26
9,77E-06
3,61
261
ATP synthase D chain, mitochondrial
Atp5h
P31399
18,8
5,7
93
49,7
7/21
1,20E-04
1,28
28
20
Actin, alpha skeletal muscle
Acta1
P68136
37,8
5,4
158
45,2
14/27
2,90E-04
1,44
44
21
F-actin-capping protein subunit alpha-2
Capza2
Q3T1K5
32,9
5,5
176
52,4
10/15
0,0000716
-1,27
-21
22
Myosin binding protein C, fast-type
Mybpc2
D3ZA38
127,2
6
148
18,1
17/25
0,0000393
-2,49
-60
23
Myosin binding protein C, fast-type
Mybpc2
D3ZA38
127,2
6
104
16,4
16/32
0,0000877
-2,16
-54
24
Myosin binding protein C, slow type
Mybpc1
Q63518
126
5,6
258
24,2
29/37
0,00104
1,2
20
25
Myosin binding protein C, slow type
Mybpc1
Q63518
126,1
5,6
154
14,3
18/27
0,000351
1,28
28
Energy balance:
TCA cycle:
Respiratory chain:
19
Structural proteins:
26
Myosin heavy chain IIB [Fragment]
Myh4
Q9JHR4
61
5,3
143
23,5
13/18
2,41E-05
3,46
246
27
Myosin-binding protein H
28
Myozenin-1
Mybph
O88599
Myoz1
D4A7U8
52,6
6
109
31,6
31,4
9,1
70
20,6
11/32
1,00E-04
-2,02
-50
8/12
0,000033
-1,28
29
Skeletal muscle-specific alpha-actinin 3
Actn3
Q8R4I6
102,9
5,2
286
-22
41,3
36/66
2,29E-05
2,01
101
30
Tropomyosin 1 alpha chain
31
Tropomyosin beta chain
Tpm1
P04692
32,7
4,6
Tpm2
P58775
32,8
4,5
225
56,3
21/46
1,45E-06
4,39
339
124
34,9
14/41
0,00000426
3,69
32
Tubulin beta-2C chain
Tubb4b
Q6P9T8
49,8
269
4,6
262
50,8
26/56
0,00416
-1,23
-19
33
Myoglobin
Mb
Q9QZ76
34
Myoglobin
Mb
Q9QZ76
17,1
8,9
163
63,6
12/27
2,83E-05
4,31
331
17,1
8,9
156
72,7
11/21
1,97E-05
5,19
419
35
Serotransferrin
Tf
36
Serotransferrin
Tf
P12346
76,3
7,8
340
36,5
24/25
1,86E-04
1,32
32
P12346
76,3
7,8
337
41,8
26/32
1,17E-04
1,45
37
Serum albumin
45
Alb
P02770
68,7
6,1
227
40,3
21/37
2,28E-05
-1,28
-22
38
Serum albumin
Alb
P02770
68,7
6,1
383
51,3
30/41
7,58E-05
-1,27
-21
Transport proteins:
Cell Signaling and Stress Proteins:
39
Alpha crystallin B chain
Cryab
P23928
19,9
6,9
141
47,7
11/28
2,50E-03
1,36
36
40
Heat shock cognate 71 kDa protein
Hspa8
P63018
70,8
5,2
226
30,2
17/20
0,00000525
-1,2
-17
41
Tripartite motif-containing protein 72
Trim72
A0JPQ4
52,8
5,8
173
36,9
14/21
0,000394
-1,25
-20
42
Voltage-dependent anion-selective channel protein 1
Vdac1
Q9Z2L0
30,6
8,6
118
34,4
8/16
3,75E-03
-1,68
-40
43
Voltage-dependent anion-selective channel protein 2
Vdac2
P81155
31,7
8,7
119
36,9
8/15
2,64E-04
-1,31
-24
44
Voltage-dependent anion-selective channel protein 2
Vdac2
P81155
31,7
8,7
100
29,2
7/14
3,14E-04
-1,29
-22