(A) ERα dependency of the pS2 promoter-LacZ reporter activity is analyzed in absence of specific H3-K9 HMTs. (B) RT-qPCR analysis was performed to document efficiency of ERα, RIZ1, and ESET siRNAs to diminish endogenous ERα, RIZ1, and ESET. (C) RT-qPCR analysis of endogenous ERα-target genes after removing H3-K9 HMTs and ERα by siRNA is shown. (D) ChIP/qPCR recruitment analysis of ERα in cells transfected with specific H3-K9 HMTs siRNAs in absence or presence of ligand (E2) is shown. (E) Analysis of pS2 promoter-LacZ reporter activity upon removing the ERα-associated coactivators CBP, pCIP, and SRC1 in cells depleted of the H3-K9 HMT RIZ1. For experiments (A) and (E), reporter plasmid and siRNAs were delivered by single-cell nuclear microinjection in MCF7 cells. In (B)-(D), siRNAs were delivered by transient transfection in MCF7 cells, and β-actin mRNA expression levels and cell transfection efficiency were used for normalization. The data in (A)-(E) are the average of three replicates, and error bars represent ± standard error mean.