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. 2016 Jan 4;12(1):e1005366. doi: 10.1371/journal.ppat.1005366

Fig 6. The c-Fos promoter is regulated via E2, AP1 and Brd4.

Fig 6

(A) ChIP analysis of three independent experiments using C33A cells transiently transfected with CRPV E2HA (grey bars) and the empty vector (control; black bars). Specific primers flanking the AP1BS at -4961 (AP1BS -4961), the canonical E2BS at position -2411 (E2BS) and the FAP1 at -454 (FAP1–454) were used to analyze respective ChIP samples. (B) Co-IP of C33A cells stably expressing CRPV E2HA, CRPV E2HA I73A, CRPV E2HA K320M/C321R and the empty vector were treated with MG132 for 16h to prevent protein degradation. Samples were precipitated using an anti HA or anti-Brd4 antibody and analyzed by Western blotting for HA and Brd4. Molecular size in kDa is indicated on the left. (C) ChIP analysis of two independent experiments. C33A cells were transiently transfected with CRPV E2HA, CRPV E2HA I73A, CRPV E2HA K320M/C321R and the empty vector and crosslinked chromatin was immunoprecipitated with HA and Brd4 antibodies. In the control sample no antibody was used. Precipitated DNA was analyzed using specific primers flanking the canonical E2BS at position -2411.