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. 2018 Sep 6;159(10):3581–3595. doi: 10.1210/en.2018-00559

Figure 2.

Figure 2.

TPA reduces recruitment of PR at specific PR target regions. (A) Easeq track visualization of PR recruitment to ACSL1, GREB1, NOTCH2, and PACSIN1 gene regions. ChIP-seq data were aligned to hg19. (B) T47D cells were treated with VC, 10 nM R5020, 1 μM TPA, and R5020+TPA for 30 minutes and subjected to ChIP assay with anti-PR and IgG as control. PR binding regions identified by ChIP-seq were validated by direct PR ChIP by quantitative real-time PCR, by use of previously published primers specific to binding regions. The data are represented as percentage input, mean ± SEM from three independent experiments. ***P < 0.0005; ****P < 0.0001. (C) T47D cells were treated with VC, 10 nM R5020, 1 μM TPA, and R5020+TPA in the presence of 1 nM E2 for 30 min and subjected to ChIP assay with anti-PR and IgG as control. The data are shown as percentage input, mean ± SEM from three independent experiments. ***P < 0.0005; ****P < 0.0001. (D) T47D cells were treated with VC, 10 nM R5020, 1 μM TPA, and R5020+TPA for 24 hours. RNA was extracted and real-time PCR analysis was performed. Primer sequences are in an online repository (26). The data are represented as fold change of VC treatment, mean ± SEM from three independent experiments. *P < 0.05; ***P < 0.0005; ****P < 0.0001. (E) Easeq track visualization of PR recruitment near candidate genes that were significantly upregulated by R5020 treatment and blocked by TPA treatment. ChIP-seq data were aligned to hg19.