Hexim1 OE alters erythroid gene expression and RNA polymerase II phosphorylation. (A) Heatmap showing genes that are differentially expressed in HEXIM1 OE compared with empty vector (EV) control. (B) Volcano plot of genes differentially expressed in HEXIM1 OE compared with EV control. (C) Gene set enrichment analyses using the ENRICHR platform of genes upregulated in EV compared with control. (D) Heatmap showing genes that are differentially expressed in HEXIM1^+/− crispr lines(+/−) compared with EV control. (E) Volcano plot of genes differentially expressed in HEXIM1^+/− compared with EV control. (F) Gene set enrichment analyses using the ENRICHR platform of genes downregulated in HEXIM1^+/− compared with control. (G). Principle component analyses of HEXIM1 OE, EV, YYFF, and ±RNA-seq studies. (H) Western blot showing Ser2 Pol II levels in EV, HEXIM1 OE, and YYFF lines (ii) with quantification (i). (I) ChIP-qPCR for Ser2 and Ser5 Pol II at the GYPA locus in HEXIM1 OE, EV, and shRNA lines. Ser2 and Ser5 Pol II occupancy at the promoter and gene body (i). Pausing index in indicated lines (ii). (J) ChIP-qPCR for Ser2 and Ser5 Pol II at the RPS19 locus in HEXIM1 OE, EV, and shRNA lines. Ser2 and Ser5 Pol II occupancy at the promoter and gene body (i). Pausing index is shown in the indicated lines (ii). *P < .05. Western blot data represent 3 independent blots of distinct cultures. ChIP-PCR data represent 2 to 3 biologic replicates.