FoxO3a regulates Puma induction in potassium deprived CGNs.

A, CGNs were maintained in high potassium (K25) or subjected to potassium withdrawal (K5). Protein extracts were collected at 8 hours and Phospho(Thr32)-FoxO3a and total FoxO3a levels were analyzed by western blot. B, CGNs were transduced with lentivirus (1 MOI) expressing shRNA directed against FoxO3a or a non-targeting control shRNA and FoxO3a protein levels were analyzed by western blot at 7 days post-infection. C, Puma mRNA levels were measured by qRT-PCR following 8 hours of potassium deprivation in CGNs transduced with FoxO3a-shRNA or non-targeting control shRNA (n = 3; *p<0.05). D, Phospho(Thr32)-FoxO3a and total FoxO3a protein was analyzed by western blot in CGNs maintained in high potassium (K25) or subjected to potassium deprivation for 8 hours in the presence or absence of either IGF-1 (200 nM), SP600125 (10 µM) or SB415286 (30 µM).