Figure 6.

FOXO3a directly binds to TGF-β and HO-1 gene promoter. (A–C) Assessment of FOXO3a binding to TGF-β and HO-1 sequence in HEK-293 T cells was performed by ChIP-PCR. The fold change values were normalized to the negative control IgG. (D–E) Wild-type and mutant vectors of TGF-β (D) and HO-1 (E) based on the two positive binding sites in ChIP were synthetized. The luciferase activities of the constructs of TGF-β and HO-1 sequence in HEK-293 T cells transfected with either FOXO3a expression plasmid or controls. All data were expressed as mean ± SEM of three independent experiments (n = 3). The differences between experimental group and NC group were analyzed using Student’s t-test. ^** P < 0.01, ^*** P < 0.001; NS, not significant versus NC group. TGF-β-WT, wild-type vectors of TGF-β; TGF-β-MUT, mutant vectors of TGF-β; HO-1-WT, wild-type vectors of HO-1; HO-1-MUT, mutant vectors of HO-1; NC, negative control.