Fig. 6.

Regulation of the Cebpa promoter by Tcfap2c and Cited2. (a) Overlapping Sp1/Sp3 and Tcfap2a/c binding sites in the Cebpa promoter. C represents overlapping Sp1/Sp3 and Tcfap2c binding site. A1 and A2 represent overlapping Sp1/Sp3 and Tcfap2a binding sites. The positions of PCR primers used for chromatin immunoprecipitation (P1 to P6) are indicated. (b) Comparison of the Tcfap2c binding sequence in the Esr1 promoter with those in the Cebpa promoters from mouse, rat, and human. (c) ChIP assays using E18.5 wild-type lungs with antibodies to Cited2, Tcfap2c, acetylated histone H3, and mouse IgG. Input lane: 1% of total chromatin was used as the PCR template. 1, 2, 3, and 4 are separate samples. (d) Re-ChIP assays by immunoprecipitating with first antibodies to Cited2 or Tcfap2c, then re-immunoprecipitating with second antibodies to Tcfap2c or Cited2. (e) Co-transfection of HeLa cells with C/EBPα-805-luc or C/EBPα-711(G/C)-luc plus Cited2, Cited2ΔCR2, Tcfap2c, CBP, and/or CBPΔCH1 in different combinations, followed by luciferase assays 24 hours after transfection. *p < 0.01. (f) Transfection of HeLa cells with si-Cited2 or si-Control for 24 hours, followed by co-transfection with C/EBPα-805-luc and Tfap2c in different combinations for 24 hours and luciferase assays. (g) Transfection of HeLa cells with si-Cited2 or si-Control for 24 hours, followed by real time RT-PCR with specific primers for human Cited2 and GAPDH.