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Fig. 4 | Breast Cancer Research

Fig. 4

From: Cholesterol biosynthesis pathway as a novel mechanism of resistance to estrogen deprivation in estrogen receptor-positive breast cancer

Fig. 4

25-HC and 27-HC enhance recruitment of ER to endogenous E-regulated genes in ER+ LTED cells. wt-MCF7 and MCF7 LTED cells were treated with E2, 25-HC and 27-HC alone or in combination with ICI182780 (ICI) for 24 hours. mRNA was extracted and quantitative RT-PCR used to measure expression of TFF1. a Data shown are representative of three independent biological experiments. Bars: ± SEM. b MCF7 LTED cells were synchronized using α-amanitin and treated with 25-HC for 45 minutes [26]. ChIP was carried out to assess the recruitment of ER to the GREB1 and TFF1 promoters, respectively. To provide evidence for an activated complex, histone deacetylase CBP recruitment was also assessed. c Effect of 27-HC on ER and CBP recruitment to the TFF1 and GREB1 promoters. Data shown are representative of three technical replicates. Bars: ± SEM. DCC Dextran-coated charcoal, E2 estradiol, ER estrogen receptor alpha, ERE estrogen response elements, HC hydroxycholesterol, LTED, long-term estrogen deprivation, wt wild type

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