Figure 2
From: PED/PEA-15 induces autophagy and mediates TGF-beta1 effect on muscle cell differentiation
Effect of TGF-beta 1 on PED/PEA-15 expression in L6 and C2C12 cells. (a) Skeletal muscle cells were stimulated with 5 ng/ml TGF-beta1 for the indicated times. Total RNA was then isolated from the cells and the levels of PED/PEA-15 mRNA were assessed by real-time RT-PCR using beta-actin as internal control. Alternatively, the cells were treated with 0.1 μM of the TGF-beta1 receptor inhibitor SB431542 or transiently transfected with 200 nM SMAD3 phosphorothioate antisense (SMAD3 AS) followed by stimulation with 5 ng/ml TGF-beta1, as described under Materials and Methods. The cells were then harvested for assessment of PED/PEA-15 mRNA levels (b) or western blotted with specific PED/PEA-15, phospho-SMAD3 (pSMAD3) and SMAD3 antibodies (c). Blots were revealed by ECL and autoradiography using beta-actin as a loading control. The autoradiograps shown are representative of four independent experiments. Bars represent the mean ±S.D. of four independent experiments. Asterisks denote statistically significant differences (*P<0.05; **P<0.01; ***P<0.001)
