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. 2012 Dec 4;23(3):340–350. doi: 10.1038/cr.2012.164

Figure 6.

Figure 6

p53 induces RoR by binding to a p53 response element in the RoR promoter. (A) Induction of RoR by p53. Cells were treated with doxo at 1 μg/ml for 24 h and then RNA was extracted for qRT-PCR. In addition, HCT-116 WT cells were transfected with vector alone or WT-p53 or mutant p53 (R175H). Values are mean ± SE (n = 3). (B) Induction of the RoR promoter luciferase activity by p53. A 1 000 bp fragment upstream of RoR was cloned into pGL3-Basic as a luciferase reporter. 293T cells were transfected with the luciferase reporter along with wild-type or mutant (R175H) p53. Luciferase assay was performed 24 h after transfection. Values are mean ± SE (n = 3). (C) Detection of p53 interaction with p53RE-1 by ChIP assay. PCR primers (RoR-ChIP-p53-1–5.1 and RoR-ChIP-p53-1–3.1) were indicated by red arrows (see Supplementary information, Figure S10B). SUMO antibody serves as a negative control, and p21 serves as a positive control.