ATAC-seq during the differentiation of BMM cells into osteoclasts

The transcription factor ETS2 was identified as a hub gene that promotes osteoclast differentiation during the progression of osteoarthritis (OA). Virtual perturbation and in vitro perturbation experiments demonstrated that knockdown of ETS2 can inhibit osteoclast differentiation. Transcriptional regulatory network analysis and combined CUT&Tag with ATAC-seq analysis results indicate that ETS2 promotes osteoclast differentiation by targeting and enhancing the expression of CEBPB. Mouse BMM cells were induced for two days with medium containing 50 ng/ml RANKL and 50 ng/ml M-CSF. ATAC-seq was used to assess the chromatin accessibility of the cells.

研究人员鉴定出转录因子ETS2为骨关节炎（osteoarthritis, OA）病程进展中促进破骨细胞分化的枢纽基因。虚拟扰动实验与体外扰动实验均证实，敲低ETS2可抑制破骨细胞分化。转录调控网络分析，以及联合靶点切割与标签整合技术（Cleavage Under Targets and Tagmentation, CUT&Tag）与转座酶可及性染色质测序（Assay for Transposase-Accessible Chromatin using sequencing, ATAC-seq）的分析结果表明，ETS2通过靶向并增强CEBPB的表达来促进破骨细胞分化。本实验采用含有50 ng/ml核因子κB受体活化因子配体（Receptor Activator of Nuclear Factor κB Ligand, RANKL）与50 ng/ml巨噬细胞集落刺激因子（Macrophage Colony-Stimulating Factor, M-CSF）的培养基，对小鼠骨髓巨噬细胞（BMM）诱导培养两天，随后通过ATAC-seq评估细胞的染色质可及性。