Figure 7
From: PED/PEA-15 induces autophagy and mediates TGF-beta1 effect on muscle cell differentiation
3-methyladenine rescues differentiation in PED/PEA-15-overexpressing L6 cells. L6 myoblasts stably overexpressing PED/PEA-15 were plated and differentiated in culture as described under Materials and Methods. Seventy two hours before the experiment, the cells were exposed to 1 mM 3-methyladenine (3MA). AVO accumulation was then assessed as outlined under Figure 4 (a). The figures shown are representative of four independent experiments. Alternatively, total RNA was obtained by the cells followed by quantitation of MyoD1, Myogenin, GLUT4 and GLUT1 mRNA levels by RT-PCR, using beta-actin as internal standard (b). Data are presented as % of the levels measured in control cells. Bars represent the mean ±S.D. of four independent experiments in which each reaction was performed in triplicate. Asterisks denote statistically significant differences (**P<0.01; ***P<0.001). (c) Brightfield images (100 × ) of L6 cells stably transfected with either the PcDNAIII-PED vector or the empty PcDNAIII vector in the absence or the presence of 3-methyadenine. The microphotographs shown are representative of three additional experiments
