Figure 6.

The number of AChR clusters induced by agrin is dependent on levels of HDAC6 activation and paxillin expression. (A) Schematic representation of the experimental time course. (B–D) After 4 d of differentiation, myotubes were treated with agrin (50 nM) for 16 h. (B) For HDAC6 inhibition conditions, WT myotubes were treated with either TubA (5 µM) or DMSO (CTL; 1 µl) at the same time as agrin. Myotubes were stained with MF-20 (in red). (C and D) Myoblasts were transfected as described in Fig. 5. For all conditions at day 5, AChRs clusters were labeled with α-BTX–A488 (in green) and myotubes with MF-20 (in red). (E) Quantifications of three independent experiments representing the total number of AChR clusters per field of view normalized to the area of all myotubes in each treated myotube condition compared with the respective control conditions (n = number of fields of 0.15 mm² counted, between 22 and 32). Means ± SEM. ***, P < 0.001; n.s, not significant; Mann-Whitney U test. (B–D) Scale bars: 100 µm.