Hemogen regulates BRG1 recruitment and chromatin accessibility at promoters and enhancer for target gene activation. (A) Volcano plot showing differential gene expression in Hemgn^−/− E14.5 fetal liver cells compared with WT littermates. Vertical dashed lines represent log₂ fold change at −0. 6 and +0. 6. Dashed horizontal line is drawn at adjusted P value <.05. Light blue dots highlight erythroid genes that were significantly downregulated upon hemogen KO (n = 4 biological replicates). (B) Immunofluorescence indicating that loss of hemogen has no obvious effect on BRG1 protein level in E14.5 fetal liver cells (left panels). Quantification of BRG1 fluorescence intensity in 30 cells (right panel). (C) Venn diagram showing global changes in BRG1 occupied peaks upon loss of hemogen in E14.5 fetal liver cells. (D) Normalized peak density plot showing BRG1 signal at hemogen binding peaks in WT or Hemgn^−/− (Mut) E14.5 fetal liver cells. (E) Heat map displaying changed BRG1 binding peaks and ATAC-seq signals between hemogen-KO and WT E14.5 fetal liver cells in 2 categories (hemogen-binding or no hemogen-binding promoters and enhancers). Each row represents a 3-kb window centered at TSS or enhancer center. N = 2 biological replicates for WT and KO ATAC-seq. (F) β-Globin locus showing ChIP-seq localization of hemogen, BRG1, GATA1, H3K27ac ChIP-seq signals, and ATAC-seq signals on the promoter and enhancers. au, arbitrary fluorescence units.