RANK ligand converts the NCoR/HDAC3 co-repressor to a PGC1β- and RNA-dependent co-activator of osteoclast gene expression [ChIP-seq]. RANK ligand converts the NCoR/HDAC3 co-repressor to a PGC1β- and RNA-dependent co-activator of osteoclast gene expression [ChIP-seq]

To investigate the genomic localization of NCoR/HDAC3/PGC1β complex and the enhancer/promoter activity in the regulation of osteoclast differentiation, we used NCoR KO or PGC1β KO bone marrow cells, non-coding RNA Dancr/Rnu12 siRNA knockdown bone marrow cells and FLAG-tagged PGC1β RNA recognition motif deletion mutant expressed RAW 264.7 cells. We then performed chromatin immunoprecipitation DNA-sequencing (ChIP-seq) for H3K27ac, NCoR, HDAC3, PGC1β, p65, Fosl2, PU.1 and FLAG-tag. Overall design: Genomic localization of NCoR/HDAC3/PGC1β and transcription factors with H3K27 acetylation depending on RANK signal

为探究NCoR/HDAC3/PGC1β复合物的基因组定位，以及其增强子/启动子活性在破骨细胞分化调控中的功能，我们使用了三类实验样本：NCoR敲除（Knockout, KO）或PGC1β敲除（KO）骨髓细胞、经非编码RNA Dancr/Rnu12小干扰RNA（small interfering RNA, siRNA）介导敲低的骨髓细胞，以及转染表达FLAG标记的PGC1β RNA识别基序缺失突变体的RAW 264.7细胞。随后我们针对H3K27ac、NCoR、HDAC3、PGC1β、p65、Fosl2、PU.1以及FLAG标签开展了染色质免疫沉淀测序（Chromatin Immunoprecipitation Sequencing, ChIP-seq）。整体实验设计：针对依赖RANK信号的H3K27乙酰化修饰，以及NCoR/HDAC3/PGC1β复合物与转录因子的基因组定位展开研究。