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. 2015 May 26;10(5):e0128239. doi: 10.1371/journal.pone.0128239

Fig 7. ChIP assay showing recruitment of ERβ2 to the HIF-1α binding site in the Twist1 and VEGF promoters.

Fig 7

In PC3 cells (A) and 22Rv1 cells (B) expressing doxycycline-regulated ERβ2 expression, this ERβ isoform binds to HRE (HIF-1α response element in Twist1 promoter) and VEGF promoter (HIF-1α response element in VEGF promoter) but not to pS2 (ERE) promoter. ChIP-qPCR results are shown with a non-specific IgG and a specific anti-M2-FLAG antibody immunoprecipitation. ERβ2 binding is enriched only in HIF-1α response element containing Twist1 and VEGF promoters but not in the pS2 promoter lacking HIF-1α response element. The graph shows the data as an increase in binding of FLAG to the Twist1 (pHRE) and VEGF promoter (mean of three separate experiments (±s.e.m.) calculated using Student’s t-test, *p≤0.028, ##p≤0.041 (PC3 cells) and **p≤0.0049, ##p≤0.0481 (22Rv1 cells).