Effect of conditional triple-knockout of Jmjd2a/Kdm4a, Jmjd2b/Kdm4b and Jmjd2c/Kdm4c or overexpression of KDM4C/JMJD2C mutants with different phase separation ability on gene expression in mouse embryonic stem cells. [ChIP-Seq]

To investigate the functions of JMJD2/KDM4 family in mouse embryonic stem cells (mESCs), we treat the the conditional triple-knockout mESCs of Jmjd2a/Kdm4a, Jmjd2b/Kdm4b and Jmjd2c/Kdm4c with 4-hydroxytamoxifen (4-OHT) for 72h. Further, to investigate whether JMJD2/KDM4 phase separation affects chromatin binding of YY1, we generated IDR-truncated JMJD2C/KDM4C (Mut) to disrupt its phase separation ability, and rescued it by by fusing two IDRs in other species (hIDR1 and hIDR2). We overexpressed those phase separation mutants in the aboved mESCs separately and TKO endogenous Jmjd2 by adding 4-OHT 72h. We then performed ChIP-seq of JMJD2A/KDM4A in WT mESC. Besides, we perform ChIP-seq of YY1, H3K27ac and H3K9me3 before and after JMJD2 TKO. Further, we perform ChIP-seq of YY1 in other 6 different cells (JMJD2C FL (Full length), JMJD2C Mut (IDR truncated), Mut-hIDR1 (rescued by hIDR1, from human HNRNPA), hIDR1, Mut-hIDR2 (rescued by hIDR2, from human FUS), hIDR2 with TKO of endogenous Jmjd2.