Fig. 4

KLF12 mediated the transcriptional repression of FOXO1. a Schematic of the location of the KLF12 binding site in the FOXO1 promoter. b hESCs were infected with the indicated adenoviruses for 8 h, transfected with FOXO1-LUC (600 ng/well), and treated with 8-Br-cAMP and MPA. After 48 h, luciferase assays were performed, and the data were plotted after normalization to Renilla luciferase activity (n = 3). **P < 0.01 and ***P < 0.001 compared with Ad-LacZ without 8-Br-cAMP or MPA; ###P < 0.001 compared with Ad-LacZ with 8-Br-cAMP and MPA. c ChIP-PCR amplification using primers against the human FOXO1 promoter region (top). PCR products were separated by agarose gel electrophoresis. Quantitative ChIP analysis was performed by real-time PCR. The results are shown as the fold enrichment over LacZ (after normalization to the input, bottom). Input (non-precipitated) chromatin was utilized as a positive control for these analyses. *P < 0.05. d ABCD assays were performed using biotinylated or non-biotinylated (competitor) double-stranded FOXO1 wild-type (WT) and mutant (MUT) oligonucleotides with whole-cell extracts from hESCs