Differential chromatine state and ER binding potentially induced by NR2F2 depletion.. Differential chromatine state and ER binding potentially induced by NR2F2 depletion.

ERα binding activity largely depends on access to binding sites on chromatin, which is facilitated in part by Pioneer Factors (PFs).We show that most binding events of NR2F2 occur together with the ERα binding sites.To explore whether NR2F2 may act as potential pioneer factor of ER, we performed a series of ChIP-seq genome wide in MCF-7. Since NR2F2 associates with chromatin prior to estrogen treatment and its depletion in MCF-7 cells did not affect ERα expression, we hypothesize NR2F2 may inhibit estrogen-dependent growth by modulating ERα recruitment. We performed ChIP-seq genome wide gainst ERα before and after NR2F2 depletion.Covalent modifications are a main chromatin property.To test whether NR2F2 favoured histone modification deposition on chromatin, we profiled ChIP-Seq of H3K4me1, H3K4me3, and H3K27ac following NR2F2 depletion in oestrogen-starved MCF-7 cells to gain comprehensive histone medication landscape. Overall design: Examination of 3 histone modifications and ER, FOXA1 binding in WT and NR2F2 KO MCF-7 cells.

雌激素受体α（ERα）的结合活性在很大程度上依赖于其对染色质结合位点的可及性，而该过程部分由先驱因子（Pioneer Factors，PFs）所介导。本研究证实，NR2F2的绝大多数结合事件均与ERα结合位点共定位。为探究NR2F2是否可作为ER的潜在先驱因子，我们在MCF-7细胞中开展了一系列全基因组染色质免疫共沉淀测序（ChIP-seq）实验。鉴于NR2F2可在雌激素处理前与染色质结合，且在MCF-7细胞中敲除NR2F2不会影响ERα的表达水平，我们推测NR2F2可能通过调控ERα的招募过程，抑制雌激素依赖的细胞增殖。我们分别在NR2F2敲除前后，针对ERα开展了全基因组ChIP-seq实验。共价修饰是染色质的核心属性之一。为验证NR2F2是否会促进组蛋白修饰在染色质上的沉积，我们在雌激素饥饿处理的MCF-7细胞中，于NR2F2敲除后对H3K4me1、H3K4me3及H3K27ac开展了ChIP-seq分析，以获取全面的组蛋白修饰图谱。实验整体设计：检测野生型（WT）与NR2F2敲除（KO）MCF-7细胞中3种组蛋白修饰以及ER、叉头框A1（FOXA1）的结合情况。