ATF4 governs the transcriptional program of erythropoiesis. (A) Experimental workflow showing the strategy used to sort the indicated cell types from the BM of fl/fl and Δ/Δ mice (n = 8-10 mice) for bulk RNA-seq, ATAC-seq, and CUT&Tag (H3K4me3). (B) Plots showing the differentially expressed genes (DEGs) identified from bulk RNA-seq analysis of fl/fl vs Δ/Δ MEP cells. (C) GO term analysis of genes identified as downregulated by bulk RNA-seq in Atf4-depleted MEP cells (vs the control group). (D) Heat map showing replication of samples of ATAC-seq from fl/fl and Δ/Δ BM MEP cell samples. (E) ATAC-seq profile and heat map showing the degree of chromatin accessibility around transcription start site (TSS) ± 3 Kbp of fl/fl and Δ/Δ MEP cells. (F) Venn diagram of ATAC-seq data showing the 29 493 different peak distributions of Atf4-deficient MEP cells. (G) H3K4me3 CUT&Tag profile and heat map showing TSS ± 3 kb. (H) Venn diagram showing the overlap between downregulated and transcriptionally repressed genes identified by ATAC-seq, H3K4me3 CUT&Tag analysis, and RNA-seq after Atf4 deletion. (I) GO term analysis of overlapping downregulated genes; only the top 11 GO terms are listed. (J) qRT-PCR analysis of the indicated genes in MEP cells sorted from the fl/fl and Δ/Δ mice (n = 3 samples). (K) Integrative genomics viewer (IGV) software was used to visually present the important genomic regions of Klf1 and Tal1 using findings from ATAC-seq and H3K4me3 CUT&Tag (shown as peaks). Data represent the mean ± SD. ∗∗∗P < .001; unpaired 2-tailed Student t test.