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. 2007 Sep 17;27(23):8228–8242. doi: 10.1128/MCB.00374-07

FIG. 7.

FIG. 7.

Mutant p53 represses the expression of TGF-βRΙΙ but not TGF-βRΙ. RT-PCR analysis of the expression of TGF-βRI/II was performed with p53-null and p53R175H-producing H1299 cells (A) or p53-null and p53R175H-producing SKOV3 cells (B). (C) Western blot analysis depicting the levels of TGF-βRI and TGF-βRΙΙ in p53-null and p53R175H-producing H1299 cells. (D) H1299 cells were transiently transfected with the TGF-βRII promoter (300 ng) along with plasmids encoding the indicated p53 mutants. A β-Gal expression plasmid was also included in all transfections, and luciferase activity was normalized to β-Gal activity. (E) SKOV3 cells were transiently transfected with the promoter of TGF-βRII together with either the p53R175H mutant, p53R175H 22-23 mutant, or wild-type p53 expression plasmid, and luciferase activity was measured and normalized to β-Gal. (F) Chromatin immunoprecipitation experiment performed on H1299 cells stably expressing the p53R175H mutant. After cross-linking of proteins to DNA, DNA was fragmented, and the p53 protein was immunoprecipitated with a specific antibody against p53 or with a nonspecific antibody against HA as a control. PCR analysis was performed on the immunoprecipitated DNA samples, using either TGF-βRII- or GAPDH-specific primers.