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Fig. 8 | BMC Biology

Fig. 8

From: The ESCRT-0 subcomplex component Hrs/Hgs is a master regulator of myogenesis via modulation of signaling and degradation pathways

Fig. 8

Inhibition of GSK3β partially rescues myoblast differentiation. a Representative Western blotting of shCT and shHrs#3 C2C12 cell extracts collected in Pro (lanes 1 and 7) and at 7, 24, 48, 72, and 96 h (lanes 2–6 and 8–12) of differentiation and probed with the anti-MHC, -GSK3β, and -phosphorylated-Tyr216 (pY216-GSK3β). GAPDH was used as a loading control. b Quantification of the pY216-GSK3β level in experiments as in a. Data are presented as ratio of pY216-GSK3β/total GSK3β and normalized to the starting point Pro condition as mean +/− SEM, n = 4 experiments. Significance was assessed using a two-way ANOVA test; *p < 0.05; **p < 0,01; ns not significant. c Representative immunofluorescence images of shCT and shHrs#3 C2C12 transduced cells treated with 3 μM of azakenpaullone (Aza) GSK3β-inhibitor or DMSO as negative control and analyzed at 120 h of differentiation. Cells were stained with anti-MHC (green) and DAPI (red). Scale bar, 200 μm. d Quantification of the myogenic index (differentiation index) corresponding to the number of nuclei present in MHC-positive structures as depicted in c. Data are presented as mean +/− SEM with 10 images per condition. n = 2 experiments

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