Fig. 3

Beclin 1, p62/SQTM1 and LAMP-1 protein levels in SMA gastrocnemius and human SMA fibroblast cell lines. Total cell lysates of gastrocnemius from P2 (A) and P5 (B) genotyped WT and mutSMA mice were submitted to western blot analysis using anti-Beclin 1, anti-p62/SQSTM1 or anti-LAMP-1, and anti-SMN antibodies. Membranes were reprobed using an antibody against α-tubulin. Graph values represent the expression of the targeted proteins vs α-tubulin at P2 and P5, and correspond to the quantification of four and three independent experiments ± SEM, respectively. Asterisks indicate significant differences using Student t test (*p < 0.05, ****p < 0.0001, no significant differences (ns) p > 0.05). C Control (unaffected) and SMA II and SMA I patient fibroblast cell lines were maintained in the presence of supplemented MEM. Forty-eight hours after plating, cell lysates were obtained and submitted to western blot analysis using anti-Beclin 1, anti-p62/SQSTM1 or anti-LAMP-1, and anti-SMN antibodies. Membranes were reprobed with an anti-α-tubulin antibody. Graph values represent the expression of the targeted proteins versus α-tubulin and correspond to the quantification of three independents experiments ± SEM. Asterisks indicate significant differences using one-way Anova with Tuckey’s multiple comparisons post-test (*p < 0.05, ***p < 0.0005, no significant differences (ns) p > 0.05)