Fig. 5.

Function of DEC1 in metformin-induced apoptosis in HeLa cells. a HeLa cells were incubated for 20 h with the indicated concentrations of metformin, after which the cell lysates were subjected to western blotting with an antibody against DEC1. ACTN was the loading control. The protein levels of DEC1 after normalization with the loading control protein ACTN are presented as fold change. b HeLa cells were incubated with 5 mM metformin with and without 10 μM MG132 for the indicated times. They were then lysed; divided into cytoplasmic, membrane, and nuclear fractions; and subjected to western blotting with antibodies against DEC1, HSP90α/β (cytoplasmic fraction), EGFR (membrane fraction) and PARP (intact: nuclear fraction; cleaved: nuclear and cytoplasmic fractions). The protein levels of cleaved DEC1, PARP, and cPARP are presented as fold change. c HeLa cells were transiently transfected with 2 μg pEGFP.DEC1 for 5 h and then were observed with fluorescence-microscopy. d HeLa cells were transiently transfected with 2 μg of pEGFP vector and pEGFP.DEC1 and incubated for 13 h with 5 mM metformin. The cell lysates were subjected to western blotting with antibodies against DEC1 and PARP. ACTN was the loading control. The protein levels of PARP and cleaved PARP (cPARP) after normalization with the loading control protein ACTN are presented as fold change. The results are representative of three independent experiments