Analysis of local glucocorticoid receptor occupancy at glucocorticoid-repressed inflammatory enhancers in Beas-2B airway epithelial cells

This study combined ChIP-seq using two different GR antibodies and stable GR knockdown in Beas-2B airway epithelial cells to determine whether glucocorticoid-mediated repression of inflammatory enhancers requires local GR occupancy Overall design: Duplicate samples of Beas-2B cells transduced with lentiviral-shRNA against GR (shGR) or a control construct (shCtrl) were treated with vehicle (ethanol), TNF (20 ng/ml), dexamethasone (100 nM) or TNF+dex for one hour prior to chromatin immunoprecipitation with one of two GR antibodies and sequencing. Duplicate Input DNA samples, representing both shCtrl and shGR groups, were included as controls.

本研究通过联用两种不同的糖皮质激素受体（GR，glucocorticoid receptor）抗体，并在Beas-2B气道上皮细胞中构建稳定GR基因敲低模型，旨在明确糖皮质激素介导的炎症增强子抑制是否依赖于局部GR结合位点的占据。实验整体设计：将经靶向GR的慢病毒短发夹RNA（shRNA）载体（shGR）或对照载体（shCtrl）转导的Beas-2B细胞设置生物学重复样本，分别以溶剂（乙醇）、肿瘤坏死因子（TNF，20 ng/ml）、地塞米松（100 nM）以及TNF联合地塞米松处理1小时，随后使用两种GR抗体中的一种进行染色质免疫共沉淀测序（ChIP-seq）。此外还纳入了分别代表shCtrl与shGR组的重复输入DNA样本作为对照。