| 描述信息 |
We established a PDHB heterozygous cell line in human embryonic stem cell HUES8. Then we performed definitive endoderm (DE) differentiation using both WT and PDHB partial deletion cell lines, and assessed gene expression and chromatin accessibility changes during this process by RNA-seq and ATAC-seq. Differentially expressed genes and regions with altered chromatin accessibility showed that in the PDHB-deleted cell line, the efficiency of endoderm differentiation was significantly inhibited. This alteration primarily occurred through changes in chromatin accessibility at BAF complex binding sites on DNA, which is a major ATP-dependent chromatin remodeling complex. And we also detected the SMAD2/3 binging by ChIP-seq.
Overall design: We performed the DE differentiation in WT and PDHB partial deletion cells. The DE differentiation medium was based on DMEM and supplemented with 100 ng/mL Activin A, 1x penicillin/streptomycin, and 0.2% bovine serum albumin. After the differentiation, we performed the RNA-seq and ATAC-seq in both cells. Besides that, we performed the SMAD2/3 ChIP-seq in WT cells. Detection was carried out after 3 days of induction using the same system. |
