Fig. 2

Increased levels of LC3-II in human SMA differentiated MNs. A Representative phase contrast and immunofluorescence images of 7-day differentiated Control and SMA human MNs, showing the MN markers HB9 (green left section), ChAT (red left section), Islet 1/2 (red right section) and beta-III-tubulin (green right section). Hoechst (blue) staining was used to identify MN nuclei. Scale bar 15 μm. B Representative immunofluorescence confocal images of 7-day differentiated human MNs using an anti-LC3 (green) antibody. Hoechst (blue) dye was used to identify nuclei. Graphs represent the mean of LC3 positive puncta per soma (above) or per neurite (1 μm) (below) and correspond to the quantification of three independent experiments ± SEM. Asterisks indicate significant differences using Student t test (****p < 0.0001). Scale bar, 15 µm. C Protein extracts of 7-day differentiated control and SMA human MNs were submitted to western blot using an anti-LC3 antibody. Membranes were reprobed with anti-α-tubulin antibody. Graph values represent the expression of LC3-II versus α-tubulin and correspond to the quantification of six independent experiments ± SEM. Asterisks indicate significant differences using Student t test (*p < 0.05)