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. 2019 Jul 18;10:3187. doi: 10.1038/s41467-019-11227-4

Fig. 6.

Fig. 6

Acute mTORC1 activation impairs HDAC4 function and limits AChR turnover. a Western blot analysis of total and phosphorylated levels of PKB/Akt and S6 in TA innervated and 3-day-denervated control (Ctrl) and iTSCmKO (iTSC) muscles, without tamoxifen treatment (−), or 6, 13, and 24 days after recombination induction (Rec). n = 3 (untreated, 6 and 13d) and 5 (28d) iTSCmKO mice. b Mass variation for TA and soleus (Sol) muscles in control and iTSCmKO mice, after 28 days of denervation, as compared to contralateral innervated muscle. n = 3 per group. c HE coloration (top panel) and fluorescent pictures of MHCIIA/X (bright/dark green) and laminin (red) immunostaining (lower panel) of TA denervated (28d) muscle from control and iTSCmKO mice. Representative of 3 independent muscles per group. Arrows, arrowhead and open arrows point to intracellular aggregates, vacuoles and abnormal giant nuclei, respectively. Scale bar, 50 (top) and 200 (bottom) µm. d Western blot analysis of HDAC4 in innervated and denervated (3d) control and iTSCmKO mice after short (sh) or long (lg) recombination induction. e, f Fluorescent pictures of HDAC4 (red) and laminin / GFP (green) of 3-day-denervated TA muscle from control and iTSCmKO mice with short or long recombination induction. Arrows point to HDAC4-positive myonuclei. Scale bar, 100 µm. Quantification in (f) gives the proportion of HDAC4-positive myonuclei. n = 5 Ctrl, 3 (short) and 5 (long) iTSCmKO muscles. g, h AChR turnover quantification for control and iTSCmKO mice with short or long recombination induction. n = 4/5/3 (In) and 7/3/3 (De) Ctrl, short and long iTSCmKO muscles. Representative pictures of “old” (green) and “new” (red) AChRs is given in (h). GFP appears in gray in iTSCmKO fibers. Scale bar, 50 µm. All values are mean ± s.e.m; two-tailed unpaired Student’s t-test (b), or one- (f) or two- (g) way ANOVA with Tukey’s post-hoc test, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. Western blot quantifications are shown in Supplementary Table 1. Source Data are provided in the Source Data File