EMBO/soda-nel huggingface dataset:
Insert L1 is a central hub for allosteric regulation of USP1 activity (2021)
Figure 3

Fig.3) USP1 (±UAF1) has higher catalytic activity on PCNA-Ub when it is loaded on DNA.

A) Schematic representation of loading and purification of PCNA-Ub on nicked circular DNA. The DNA-loaded PCNA-Ub elutes in the void of the SEC column and this sample is collected and used for studying activity of USP1 on DNA-loaded PCNA-Ub.

B) Michaelis-Menten analysis of USP1+UAF1 with and without DNA (65bp dsDNA) shows that DNA binding alone has no effect on USP1 activity (n=2, technical replicates (example shown here from 1 of 2 biological replicates)).

C) Coomassie stained gel of in vitro activity assay showing increased activity of USP1-UAF1 on DNA-loaded PCNA-Ub compared to PCNA-Ub (-DNA) and PCNA-Ub (+DNA, +RFC and not ATP).

D) Quantification of gel-based activity assays showing enhanced activity of USP1+UAF1 on DNA-loaded PCNA-Ub compared to free PCNA-Ub (n=3, technical replicates (example shown here from 1 of 2 biological replicates)).

E) Quantification of gel-based activity assays showing enhanced activity of USP1 on DNA-loaded PCNA-Ub compared to free PCNA-Ub (n=3, technical replicates (example shown here from 1 of 2 biological replicates)).
Insert L1 is a central hub for allosteric regulation of USP1 activity (2021)
Figure 4

Fig.4) Modelling the enhanced USP1 activity on DNA-loaded PCNA-Ub.

A) Gel based quantification of USP1-UAF1 activity on PCNA-Ub (DNA loaded and free) for three different concentration of enzyme and substrate.

B) FP based activity assays on free PCNA-Ub (three concentrations of USP1 and PCNA-Ub) and activity of USP1 on increasing concentrations of Ub-Rho.

C) Kinetic model of USP1-UAF1 activity on PCNA-Ub and DNA-loaded PCNA-Ub (Ub-Rho not shown here, see EV 4c for full model), constants with the same colour were linked during the fitting and they share same values.
Insert L1 is a central hub for allosteric regulation of USP1 activity (2021)
Figure 5

Fig.5) Insert L1 mediated DNA and PCNA interactions are crucial for increase in USP1 activity on DNA loaded PCNA-Ub

A) Comparison of USP1WT and deletion mutants (+UAF1) for activity on DNA-loaded PCNA-Ub (solid lines) and free PCNA-Ub (dashed lines) shows no increase in USP1ΔL1 activity on the loaded substrate. Left panel: Quantification of gel-based activity assays showing percentage of cleaved PCNA-Ub at the mentioned time points (n=2, biological replicates), Right panel: Quantification of the activation fold observed in USP1WT and mutants on DNA-loaded PCNA-Ub versus free PCNA-Ub (n=2, biological replicates).

B) Multiple sequence alignment of USP1 insert L1 which highlights the DNA binding region and the conservation of the positively charged residues across species. Three groups of positively charged residues (***) are mutated separately and tested for binding to double stranded DNA (65bp) using SPR which shows the importance of this region for DNA binding of USP1.

C) Comparison of USP1DM1, USP1PIP1 and USP1PIP1+DM1 (+UAF1) activity on DNA-loaded PCNA-Ub (solid lines) and free PCNA-Ub (dashed lines) shows reduced increase in activity of these mutants compared to USP1WT. Left panel: Quantification of gel based activity assays showing percentage of cleaved PCNA-Ub at the mentioned time points (n=2, technical replicates), Right panel: Quantification of the activation fold observed in USP1WT, USP1PIP1, USP1DM1 and USP1PIP1+DM1 on DNA-loaded PCNA-Ub versus free PCNA-Ub (n=2, technical replicates).

D) EMSA based DNA binding experiment shows that USP1PIP1 and USP1WT has similar DNA binding capability while USP1DM1 has lost its ability to bind DNA.

E) Schematic model for the role of Insert L1 in USP1 activity regulation by UAF1 and DNA loaded PCNA-Ub.

Table.1 Kinetic analysis of USP1WT and deletion mutants using the Michaelis-Menten equation.

kcat

(sec-1)

KM

(µM)

kcat/KM

(sec-1 µM-1)

Table.2 KinTek modelling of USP1 (±UAF1) activity on three substrates i.e. Ub-Rho, PCNA-Ub and DNA-loaded PCNA-Ub

kcat

(kcut)*#

* Constants with the same symbol (α, β, γ, δ) were linked during the fitting and they share same values

# Although kcut mathematically is not equivalent to Michaelis-Menten catalytic rate constant kcat the difference in the values between them in this model is neglectable in comparison to measurement and fitting errors.

Expanded View Figure legends
TCF20 dysfunction leads to cortical neurogenesis defects and autistic-like behaviors in mice (2020)
Figure 1

Figure 1 TCF20 is expressed in the developing cerebral cortex and neural stem cells

(a) Immunostaining of TCF20, NESTIN, and merged with DAPI in E13 and E16 embryonic brain sections. TCF20 is widely expressed in the brain.

(b) Costaining with the NSC markers NESTIN, SOX2, and PAX6 reveals that TCF20 is expressed in NSCs.

(c) Absolute quantitative PCR of TCF20 in different developmental stages from E12 to P0. n=4 independently repeated tests.

(d) Western blotting results show the changes in TCF20 protein expression level during brain development.

(e) Statistical analysis of normalized band intensity in the western blot results. n= 4 samples for each group.

(f) In utero electroporation of TCF20 shRNAs in the E13 cerebral cortex led to abnormal cell distribution. These electroporated embryos were collected at E16.

(g) Statistical analysis of the distributions of GFP-positive cells in different brain regions. The percentage of GFP-positive cells in the CP (cortical plate), IZ (intermediate zone), and VZ/SVZ (ventricle zone and subventricle zone) was analyzed. n=7 brains.

Data information: Error bars represent the means ± S.E.M. Two tailed unpaired T-tests were used to analyze the data, n.s. (no significant difference), p<0.01(**), p<0.001(***). Scale bar in (A) e13, 50 μm, e16 100 μm; (E) 50μm; (F) 50 μm.
TCF20 dysfunction leads to cortical neurogenesis defects and autistic-like behaviors in mice (2020)
Figure 2

Figure 2 TCF20 deficits result in abnormal cell proliferation

(a) A diagram of the BrdU labeling time point in electroporation experiments.

(b) Immunostaining of BrdU. White arrows represent BrdU GFP double-positive cells. Scale bar: 50 μm.

(c) Statistical analysis of 2-hour BrdU labeling experiments. n=6 brains

(d) Western blotting analysis indicated an increase in the proliferation marker Sox2 in TCF20 depleted NSCs.

(e) Immunostaining with pax6 in E16 electroporated embryo brains. White arrows represent PAX6, GFP double-positive cells. 1,2 are 2x enlarged as the dashed boxes showed. Scale bar: 50 μm.

(f) Statistical analysis of the percentage of PAX6+GFP+ cells among the total GFP+ cells. n=3 brains for each group.

(g) Immunostaining with sox2 and pax6 in E16 electroporated embryo brains. White arrows represent GFP, sox2 double positive cells; Red arrows represent GFP, TBR2 double positive cells. area. Scale bar: 50 μm.

(h) Statistical analysis of the percentage of SOX2+GFP+ cells among the total GFP+ cells. n=8 brains for each group.

(i) Statistical analysis of the percentage of TBR2+GFP+ cells among the total GFP+ cells. n=5 brains for each group.

Data information: Error bars represent the means ± S.E.M. Two tailed unpaired T-tests were used to analyze the data, n.s. (no significant difference), p<0.05(*), p<0.01(**), p<0.001(***).
TCF20 dysfunction leads to cortical neurogenesis defects and autistic-like behaviors in mice (2020)
Figure 3

Figure 3 TCF20 depletion impairs neuronal differentiation

(a) SATB2 staining revealed a reduction in SATB2 and GFP colocalization when TCF20 was depleted. White arrows represent SATB2, GFP double-positive cells.

(b) Statistical analysis of the SATB2 GFP double-positive cell ratio in GFP positive cells. n=3 brains.

(c) Staining with CTIP2 and TBR1 antibodies shows TCF20 knockdown decreases the colocalization of CTIP2 and TBR1 with GFP in E13.5-E16.5 brains. White arrows represent CTIP2, GFP double-positive cells. White arrow heads represent TBR1, GFP double-positive cells.

(d, e) Statistical analysis of CTIP2, GFP and TBR1, GFP double-positive cell ratio in GFP positive cells. n=3 brains.

(f) A diagram of the BrdU labeling time point in electroporation experiments.

(g) Immunostaining of BrdU. White arrows represent BrdU GFP double-positive cells.

(h) Percentage of BrdU and GFP double-positive cells in GFP-positive cells n=3 brains for each group

Data information: Error bars represent the means ± S.E.M. Two tailed unpaired T-tests were used to analyze the data, n.s. (no significant difference), p<0.05(*), p<0.01(**), p<0.001(***). Scale bar: (b,f) 25 μm; (d) 50 μm
TCF20 dysfunction leads to cortical neurogenesis defects and autistic-like behaviors in mice (2020)
Figure 4

Figure 4 Deletion of TCF20 impairs neurogenesis

(a) A diagram of the construction of TCF20 knockout mice. Two guide RNAs targeted the ends of TCF20 exon 2. With the help of cas9, TCF20 exon 2 was deleted from the genome.

(b) Western blotting analysis of TCF20 in TCF20 WT, HET, KO mice reveals that TCF20 was deleted.

(c) Statistical analysis of the normalized band intensity of TCF20. n=3 samples for each genotype.

(d) RT-qPCR analysis of TCF20 in TCF20 WT, HET, and KO mice reveals that TCF20 was deleted. n=4 samples.

(e) Immunostaining of E16 WT and KO mouse brain slices revealed that TCF20 was successfully knocked out. Scale bar: 50 μm.

(f) Statistical analysis of the normalized fluorescence intensity of TCF20. n=6 samples for each genotype.

(g) Western blotting analysis of neural proliferation and differentiation markers in WT HET and KO mice showed that TCF20 deficits promote cell proliferation and inhibit cell differentiation.

(h) Immunostaining of different cortical layer markers TBR1, CTIP2, and SATB2, White dash line represents the positive area. Scale bar: 50 μm.

(i-k) Statistical analysis of different cells layers in TCF20 WT, HET and KO mice. n=7 brains of each group.

(l) TCF20 deletion impairs neurogenesis and results in abnormal cell distribution. Scale bar: 50 μm.

(m) Statistical analysis of GFP+ distribution in different zones. n=3 brains for each genotype.

Data information: Error bars represent the means ± S.E.M. Two tailed unpaired T-tests were used to analyze the data, n.s. (no significant difference), p<0.05(*), p<0.01(**), p<0.001(***).
TCF20 dysfunction leads to cortical neurogenesis defects and autistic-like behaviors in mice (2020)
Figure 6

Figure 6 TCF20 regulates neurogenesis through TDG

(a) GO analysis of 215 significantly downregulated genes (Log2 fold change < -1, -P value < 0.05)

(b) GO analysis of 197 significantly upregulated genes (Log2 fold change > 1, -P value < 0.05)

(c) Heat map of significantly different genes.

(d) TDG is significantly decreased after TCF20 deletion.

(e) Volcano map showing that TDG is significantly decreased.

(f, g) Electroporation of GFP at E13.5 to E16.5 in TCF20 KO mice impairs cell distribution. n=3 brains. Scalebar 25 μm

(h) Chip-qPCR of TCF20 binding to the TDG promoter. n= 4 biological repeats.

(i) Combining RNA-seq and TDG ChIP-seq data, TCF-4 is a downstream target gene. Data information: Error bars represent the means ± S.E.M. Two tailed unpaired T-tests were used to analyze the data, n.s. (no significant difference), p<0.05(*), p<0.01(**).
TCF20 dysfunction leads to cortical neurogenesis defects and autistic-like behaviors in mice (2020)
Figure 7

Figure 7 T cell factor-4 is responsible for TDG controlling neural differentiation

(a) Diagram of the TCF-4 promoter region.

(b) TDG ChIP-qPCR revealed that TDG mainly binds to TCF-4 CpG islands. Seven regions (from -5 kbp to -200 bp) of the promoter were analyzed. -500 bp and -200 bp regions were located in the CpG island of the TCF-4 promoter. (n=4 biological repeats).

(c-f) MeDIP-qPCR showed that 5fC, 5caC, and 5mC are increased in the TCF-4 promoter, while 5hmC is decreased when TCF20 is insufficient. The MeDIP assay measure the region -500 bp from the TCF-4 promoter. (n=4 biological repeats)

(g) NSCs were isolated from C57B6/J E13.5 embryonic brains and then were infected with a lentivirus containing a TCF20 shRNA or with a control plasmids. Immunostaining of 5fC shows that TCF20 depletion increased 5fC levels. White dashed lines show the cells infected with lentivirus. Scalebar 25 μm

(h)Statistical analysis of 5fC fluorescence level. n=20 cells, and P<0.0001

(i) Immunostaining of 5caC shows that TCF20 depletion increased 5caC level. White dashed lines show the cells infected with lentivirus. Scalebar 25 μm.

(j) Statistical analysis of the 5caC fluorescence level. n=20 cells, and P<0.0001.

(k, l) Overexpression of TDG and TCF-4 rescued the TCF20 knockdown phenotype in brains; n=3 brains. Scalebar 50 μm

Data information: Error bars represent the means ± S.E.M. Two tailed unpaired T-tests were used to analyze the data, n.s. (no significant difference), p<0.05(*), p<0.01(**).

and p<0.001(***).
Contraceptive progestins with androgenic properties stimulate breast epithelial cell proliferation (2021)
Figure 1

Figure 1- Ex vivo stimulation of murine mammary organoids with different PR agonists.

A Experimental scheme showing the preparation of organoids from mouse mammary glands and ex vivo stimulation with different compounds.

B PCA plot of global gene expression profiles from ethanol (CTRL) and R5020-treated murine mammary organoids.

C Unsupervised hierarchical clustering of the 500 most variable genes across samples. Normalized expression levels were scaled to Z-scores for each gene.

D Volcano plot showing differentially expressed genes (adj. P.Val < 0.05) between ethanol- and R5020-treated mouse mammary organoids, n=3. Genes with log2(FC) > 2 are highlighted in red, genes with log2FC < -2 in blue. Names of genes with -log10 (adj.P.Val) > 10 and abs(log2(FC)) > 2 are indicated.

E GSEA showing enrichment of pathways differentially regulated in mouse mammary organoids upon R5020 stimulation, total number of genes = 14,244. The red dashed line indicates NES: Normalized Enrichment Score.

F Bar graphs showing Rankl and Wnt4 transcript levels, relative to 36B4 mRNA expression in mammary organoids derived from C57Bl/6 females 6 hours after stimulation with 20 nM R5020 (n=12), DSG (n=6), GSN (n=6), LNG (n=6), CMA (n=6), CPA (n=6), or DSP (n=6).

G Bar graphs showing relative Rankl and Wnt4 transcript levels normalized to 36B4 mRNA expression in mammary organoids derived from Balb6/C females 24 hours after stimulation with different progestins, n=3 independent experiments.

H, I Bar graphs showing relative Rankl and Wnt4 transcript levels normalized to 36B4 expression in C57Bl6-derived mammary organoids treated for 6 hours with R5020 (H) or LNG (I) and either 10 μM or 100 μM enzalutamide, n=3 independent experiments.

Data information F-I: Data are shown as means ± SD. P-values were estimated using a random-intercept linear model applied to ΔCt values followed by Dunnett test. * P <0.05, ** P < 0.01, *** P < 0.001, n.s: not significant.
Contraceptive progestins with androgenic properties stimulate breast epithelial cell proliferation (2021)
Figure 3

Figure 3- Human breast epithelial cell xenografts are hormone-responsive.

A Scheme of the mouse intraductal xenograft (MIND) approach. Freshly isolated human breast tissue is dissociated to single cells, which are lentivirally transduced and injected into the milk ducts via the teat. In vivo growth is monitored by weekly luminescence measurements. At endpoint, xenografted mammary glands and blood are collected for further analyses.

B Spaghetti plot showing fold change radiance over time, individual xenografted mammary glands (light grey), averaged control samples from individual xenografts (dark grey), average values of control samples from all xenografts (black, red dots), n=12-224 xenografted glands per time point.

C Fluorescence stereo micrograph of inguinal mammary gland 10 weeks after injection of GFP labelled HBECs. Dashed lines highlight the perimeter of the mammary fat pad. Scale bar, 3 mm.

D Stereo micrographs of whole mounted mammary glands 12 weeks after injection of HBECs. Scale bar, 2 mm. Inset, arrows point to mouse ducts distended by the presence of HBECs. Scale bar, 0.8 mm.

E H&E stained section of xenografted gland 83 days after injection showing HBECs that distend a mouse duct (arrow), small mouse ducts are shown (small arrow heads). Scale bar, 0.1 mm.

F Fluorescence micrograph of histological section of an intraductally xenografted mammary gland stained with DAPI (top) and immunofluorescence with anti hCK7 (bottom). Arrows point to mouse ducts devoid of human cells. Scale bar, 0.1 mm.

G, H IHC for hCK19, in brown, of cross-sectioned "humanized" mouse milk duct mammary glands 106 days after injection. Arrows point to mouse ducts colonized by human cells. Scale bars, 100 μm (G) and 150 μm (H), respectively.

I Representative co-IF micrographs of a cross-sectioned "humanized" mouse milk duct 5 months after intraductal injection for AR, ERα and PR, in red, and human E-CAD, in green. Scale bar, 50 μm.

J IHC for hCK7, in brown, of cross-sectioned "humanized" mouse milk duct mammary glands at day 20 of pregnancy. Arrow heads point to mouse alveoli, stars highlight lumina filled with secretions. Scale bar, 100 μm.

K Bar graphs showing the threshold cycles of semi quantitative RT-PCR with human-specific primers on RNA from virgin (n=4) and pregnant (n=3) mice xenografted with human cells.
Contraceptive progestins with androgenic properties stimulate breast epithelial cell proliferation (2021)
Figure 7

Figure 7- AR is required for levonorgestrel-driven proliferation of xenografted HBECs.

A-C Graphs showing in vivo growth of HBECs from different donors as measured by radiance after treatment with LNG, ENZ, or LNG and ENZ, means of radiance in individual glands ± SEM, n= 8-10 per treatment, Wilcoxon matched-pairs test. Right: Dot plot showing plasma levels of progesterone (P), testosterone (T) and LNG in individual xenografted animals.

D Graph showing in vivo growth of HBECs from a 20-year-old patient transduced with either sh scramble or shAR and treated with LNG. Points show means of radiance in individual glands ± SEM; n= 8-10 per treatment. Wilcoxon matched-pairs test.

E Representative micrographs showing co-IF with anti-GFP (red), and anti-AR (green) antibodies on histological sections from glands xenografted with HBECs transduced either with sh scramble or shAR-expressing lentivirus. Scale bar, 50 μm.

F Violin plot showing the percentage of AR- and GFP-double+ cells of total GFP+ cells in sh scramble (n=6) and shAR (n=12) conditions, dots represent individual sectors counted, median (red). Statistical significance was assessed by fitting a generalized linear mixed model with gamma distributions using CTRL as reference. Red line shows median.

Data information: * P < 0.05, ** P < 0.01, *** P < 0.001, n.s: not significant.
Succinate induces skeletal muscle fiber remodeling via SUNCR1 Signaling Pathway (2019)
Figure 1

FIGURE 1. Effects of succinate on growth performance and serum concentration in mice.

(A-G) Male C57BL/6J mice were fed with chow diet supplemented with 0, 0.5%, 1% SUC for 8 weeks.

(A-E) (A) Serum SUA level, (B) body weight gain, (C) fat and (D) lean mass and (E) gastrocnemius index.

(F-G) (F) Gastrointestinal muscle fiber immunofluorescent laminin staining and (G) frequency histogram of fiber cross-sectional area. Scale bar in (F) represents 100 µm.

Data information: Results are presented as mean ± SEM (n=6-8). Different letters between bars mean P ≤ 0.05 in one way ANOVA analyses followed by post hoc Tukey's tests. *: significant difference (P ≤ 0.05) between 0.5% SUC and control group by non-paired Student's t test. #: significant difference (P ≤ 0.05) between 1% SUC and control group by non-paired Student's t test.
Succinate induces skeletal muscle fiber remodeling via SUNCR1 Signaling Pathway (2019)
Figure 3

FIGURE 3. Effects of succinate on MyHC expression in mice.

(A-D) Male C57BL/6J mice were fed with chow diet supplemented with 0, 0.5%, 1% SUC for 8 weeks.

(A) mRNA expression of MyHC I, MyHC IIa, PGC-1α, myoglobin, TnnT1 MyHC IIb, MyHC IIx and TnnT3 in the gastrocnemius muscle (n=5-6).

(B) Immunoblots and quantification of MyHC-I, IIa and IIb protein expression in gastrocnemius (n=3-4).

(C-D) Representative images and quantification of Laminin (green), MyHC-I and IIb immunofluorescent staining (red) in gastrocnemius (n=3). Scale bar in (C) represents 100 µm.

Data information: Results are presented as mean ± SEM. Different letters between bars mean P ≤ 0.05 in one-way ANOVA analyses followed by post hoc Tukey's tests.
Succinate induces skeletal muscle fiber remodeling via SUNCR1 Signaling Pathway (2019)
Figure 4

FIGURE 4. Succinate promotes skeletal muscle mitochondrial biosynthesis and aerobic oxidation in mice.

(A-I) Male C57BL/6J mice were fed with chow diet supplemented with 0 and1% SUC for 6 weeks.

(A-D) The O2 consumption (VO2) (A, B) and respiratory exchange ratio (RER) (C, D)

(E-H) Serum concentration of (E) NEFA in whole blood. The enzymes activity of (F) SDH, (G) HK, and (H) LDH in gastrocnemius.

(I-J) Immunoblots and quantification of of p-AMPK，PGC-1α and myoglobin in gastrocnemius. The same lysates were used for the detection of PGC1α (100 kDa, Fig. 4I), myoglobin (17 kDa, Fig. 4I), myosin heavy chain (180 kDa, Fig. 3B) and tubulin (48 kDa, shared in both Fig. 3B and 4I).

(K) Quantification of mitochondrial and electron transport chain (ETC) related genes expression in gastrocnemius.

(L) OCRs were measured under basal condition in gastrocnemius.

Data information: Results are presented as mean ± SEM (n=4-6). Different letters between bars mean P ≤ 0.05 in one-way ANOVA analyses followed by post hoc Tukey's tests. *P ≤ 0.05 and **P ≤ 0.01 by non-paired Student's t test.
Succinate induces skeletal muscle fiber remodeling via SUNCR1 Signaling Pathway (2019)
Figure 5

FIGURE 5. Effects of succinate on MyHC expression, mitochondria biosynthesis and metabolism in C2C12 cells.

(A-J) C2C12 cells were treated with 0, 0.5 and 2 mM SUC for 48 h.

(A-B) Representative images and quantification of MyHC-I and IIb immunofluorescent staining (green) in C2C12 cells (n=16).

(C-E) The enzymes activity of (C) SDH, (D) LDH, and (E) lactate production in C2C12 cells.

(F) Quantification of mitochondrial DNA contents in C2C12 cells.

(G-J) (G) Mitochondrial electron microscopy showed the (H) mitochondrial density, (I) mitochondrial coverage, and (J) average mitochondrial area in C2C12 cell. Scale bar in (A) represents 50 μm, scale bar in (G) represents 0.5 μm.

Data information: Results are presented as mean ± SEM (n=6-8). Different letters between bars mean P ≤ 0.05 in one-way ANOVA analyses followed by post hoc Tukey's tests. *P ≤ 0.05 by non-paired Student's t test.
Succinate induces skeletal muscle fiber remodeling via SUNCR1 Signaling Pathway (2019)
Figure 6

FIGURE 6. SUNCR1 is required for succinate to induce the fiber type transition in myotubes.

(A-B) SUNCR1 protein expression in the gastrocnemius from sedentary or post-running mice (n=4).

(C) The mRNA level of SUNCR1 in gastrocnemius and soleus (n=7-8).

(D) [Ca2+]i in C2C12 cells treated with 0 or 2 mM SUC (n=18-20).

(E-G) NFAT protein expression in nucleus and cytoplasm of gastrocnemius 0.5 h or 3 h after i. p. injection of 15 mg/kg succinate in C57BL/6J mice (n=4).

(H) SUNCR1 protein expression in C2C12 cells transfected with vector or siSUNCR1 (n=3).

(I-L) (I) [Ca2+]i, and enzymes activity (n=9-10)of (J) HK, (K) LDH, and (L) SDH in vector or siSUNCR1 transfected C2C12 cells treated with 0 or 2 mM SUC (n=5-6).

(M-O) Representative images and quantification of MyHC-I and IIb immunofluorescent staining (green) in C2C12 cells (n=3). Scale bar in (M) represents 50 μm.

Data information: Results are presented as mean ± SEM. *P ≤ 0.05 by non-paired Student's t test.