FIGURE 6.

C/EBPα binds to the steap4 promoter during feeding and binding decreases during fasting. A, ChIP assays were performed using livers from mice that were either fed or fasted to detect the presence of C/EBPα and STAT3 at the steap4 promoter or the control β-globin locus. Band intensities were measured and graphed. B, C/EBPα protein levels in the liver were detected by Western blot (n = 3); c/ebpα mRNA levels were measured by qPCR analysis (n = 8). C, Western blot (C/EBPα, GFP, and actin) and qPCR (STEAP4) results from livers of mice overexpressing either GFP (control) or C/EBPα (n = 6–10) (D). HepG2 cells were transfected with pGL3-steap4 with or without co-transfected C/EBPα, and luciferase activity was measured and normalized to total protein levels (n = 4). E, ChIP assays were performed to detect the presence of C/EBPα and STAT3 at the steap4 and socs3 promoters in livers from mice treated with saline or IL-6; band intensities were measured and graphed. F, C/EBPα and actin protein levels in saline- and IL-6-treated livers were detected by Western blot (n = 4). *, p < 0.05; **, p < 0.001; NS, not significant; Ab, antibody.