Fig. 4

Changes of mTOR protein level and phosphorylation at Ser2448 in protein extracts of SMA tissues and human SMA fibroblast cell lines. A Total cell lysates of P2 and P5 gastrocnemius from WT and mutSMA genotyped mice were submitted to western blot analysis using anti-mTOR, anti-phospho-mTOR(Ser2448) and anti-SMN antibodies. Membranes were reprobed using an anti-α-tubulin antibody. Graph values represent the expression of mTOR or phospho-mTOR(Ser2448) (p-mTOR) versus α-tubulin and correspond to the quantification of at least five independent experiments ± SEM. Asterisks indicate significant differences using Student t test (*p < 0.05; ****p < 0.0001; non statistic (ns) p > 0.05). B Protein extracts from 48 h cultured control (unaffected) and SMA II and SMA I patient fibroblast cell lines were submitted to western blot using anti-mTOR or anti-phospho-mTOR(Ser2448) and anti-SMN antibodies. Membranes were reprobed with an anti-α-tubulin antibody. Graph values represent the expression of mTOR or p-mTOR versus α-tubulin and correspond to the quantification of five independent experiments ± SEM. Asterisks indicate significant differences using one-way Anova with Dunnett’s multiple comparisons post-test (***p < 0.001; ****p < 0.0001)