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. Author manuscript; available in PMC: 2014 Aug 27.
Published in final edited form as: Oncogene. 2013 Apr 1;33(9):1101–1112. doi: 10.1038/onc.2013.69

Figure 5. 15-PGDH-derived 15-keto-PGE2 enhances PPARγ association with the p21 promoter and induces p21 expression in HCC cells.

Figure 5

Figure 5

A. The effect of 15-PGDH and 15-keto-PGE2 on p21 promoter activity. a p21 promoter luciferase reporter activity in Huh7 cells with altered expression of 15-PGDH (**p < 0.01). b p21 promoter luciferase reporter activity in Huh7 stable cell lines with co-transfection of PGR2. c p21 promoter luciferase reporter activity in Huh7 stable cell lines treated with 10µM GW9662. d 15-keto-PGE2 (10µM) enhanced p21 promoter luciferase reporter activity in wild type Huh7 cells; the effect was abolished by PGR2 overexpression or by GW9662 treatment (10µM).

B. PPARγ CHIP-PCR assay. a PPARγ association with p21 promoter in Huh7 stable cells with or without PGR2 overexpression. b 15-keto-PGE2 (10µM) enhanced PPARγ binding to the p21 promoter in wild type Huh7 cells and the effect was abolished by PGR2 overexpression or GW9662 treatment (10µM). IgG CHIP was used as the negative control. p21 promoter PCR product was used as the input.

C. PPARγ CHIP-real-time PCR assay. a PPARγ association with p21 promoter in Huh7 stable cells with or without PGR2 overexpression. b 15-keto-PGE2 (10µM) enhanced PPARγ binding to the p21 promoter in wild type Huh7 cells and the effect was abolished by GW9662 treatment (10µM). IgG CHIP was used as the negative control.

D. Effect of the PPARγ agonist ciglitazone (10µM) in wild type Huh7 cells. a PPARγ CHIP-real-time PCR assay. b PPRE luciferase reporter activity assay.

E. Western blotting for p21 and phosphorylated p21 in Huh7 cells with indicated transfections.