FoxO1 mediates PC2 production.
A–D, N43-5 cells were transfected with either 4 μg of GFP cDNA or FoxO1 cDNA (n = 6/group) and were collected 48 h post-transfection. A, FoxO1 protein. B, PC1 protein. C, PC2 protein. D, Sirt1 protein levels measured by Western blotting analysis. These results were repeated in an independent experiment with a similar response (n = 6/group). E–H, in another experiment, AtT20 cells were transfected as just described and were treated with Ex-527 (50 μm) or with vehicle control for 6 h and collected 48 h post-transfection. E, PC2 protein (n = 12/group); F, FoxO1 protein (n = 6/group); G, acFoxO1 protein (n = 6/group); H, Sirt1 (n = 9 GFP-vehicle (GV), 13 FoxO1-vehicle (FV), and 7 FoxO1-Ex527 (FE)); I, PC2 protein (n = 9 GFP-vehicle (GV), 12 FoxO1-vehicle (FV), and 12 FoxO1-Ex527 (FE), and protein levels were measured by Western blotting analysis. Data are mean ± S.E. *, p value< 0.05; **, p value < 0.01; ***, p value < 0.001; ****, p value < 0.0001.