Figure 9

Functional consequences of PED/PEA-15 overexpression in skeletal muscle of transgenic mice. (a) Tibialis anterior muscles from PED/PEA-15 transgenic mice and from their non-transgenic littermates (WT) were dissected, solubilized and 100 μg protein samples were analyzed by western blotting using LC3-I, LC3-II, phospho-tyr307-PP2A (P-PP2A), PP2A, phospho-ser256-FoxO1 (p-FoxO1) and FoxO1 antibodies. Beta actin was used as loading control. Blots were revealed by ECL and autoradiography. The autoradiograph shown is representative of studies performed with n=8 mice/group. Alternatively (b), the muscle was used for obtaining total RNA preparation. MyoD1 and Myogenin mRNA was subsequently assessed by real-time RT-PCR analysis, using beta-actin as internal standard. Bars represent the mean±S.D. of four independent experiments each performed in triplicate. (c) Travel distance was compared in transgenic and non-transgenic littermates (16 animals/group) enabled to explore the open-field appartus for 5 min as described under Materials and methods. Bars represent the mean±S.D. of four independent studies each involving four mice/group. Asterisks denote statistically significant differences (^*P<0.05; ^***P<0.001)