Cohesin regulates alternative splicing alone or with BRD4

By targeted depletion of Cohesin (RAD21 subunit) or BRD4 alone or together, we have established that cohesin alone or with BRD4 regulates the alternative splicing of a large subset of genes under both physiological and heat shock conditions. We examined by RNA-seq the effect on gene expression and alternative splicing of cohesin and BRD4 deletion alone or together under physiological and heat shock conditions. Targeted degradation of RAD21 was mediated by an auxin-inducible degron (AID) system (HCT116RmAC cells, RAD21 alleles are fused with a minimal AID tag) and the targeted depletion of BRD4 in HCT116RmAC cells was achieved by treating with MZ1, a proteolysis targeted chimera (PROTAC), under conditions leading to selective degradation of BRD4.