Stag2 perturbation leads to preferential loss of cohesin binding at differentiation promoters with strong Etv6 prebinding. (A) Representation of the erythroid perturbation tiers; differences of Rad21 binding in Luc-Ery and shS2-Ery cells (Rad21-Ery − shS2-Ery) in the indicated erythroid perturbation tiers. Box plots to the left of 0 indicate increased binding following Stag2 knockdown and to the right indicate increased binding in control knockdown cells with normal erythroid differentiation. (B) Differential H3K27ac binding as in panel A. (C) Differential significant interaction frequencies at the promoters of the same regions as in panel A. (D) Overlaps between annotated promoters from the indicated perturbation tiers and differentially expressed genes from Figure 4F. (E) Enrichment of the specified hTF at the perturbation tiers in wild-type HPC. (F) Enrichment of Etv6 at the indicated regions. (G) Immunoblotting for total protein expression of Etv6, Stag2, and actin in the indicated conditions. (H) Immunoblotting for protein expression of Etv6, Stag2 H3, and actin in different cellular fractions in the indicated conditions. (I) Immunoblotting for protein expression of Etv6, Stag2 H2AX, and actin in different cellular fractions in the indicated conditions. (J) ChIP-qPCR of Etv6 binding at Klf1, Epor, and Cxcr4 promoter regions in the indicated cellular states. Shown are results from 3 experimental replicates.