Regulation of gene expression by BCL11A during in vivo erythropoiesis. CD34⁺ HSPCs were electroporated with RNP targeting BCL11A^E and transplanted into NBSGW mice. Untreated CD34⁺ HSPCs were transplanted as a negative control. After 16 weeks, hCD235⁺ donor erythroblasts were isolated from recipient BM and analyzed by RNA-seq. (A) Volcano plot illustrating gene expression changes in BCL11A^E-disrupted erythroblasts. Dashed lines represent the 0.05 FDR statistical significance cutoff (y-axis) and log₂ fold change of 1 or −1 on the x-axis. (B) Overlap of DEGs (fold change > 2 and FDR < 0.05) between BCL11A^E-disrupted vs control erythroblasts generated in vivo and BCL11A^E-disrupted vs control basophilic erythroblasts generated in vitro. (C) Scatterplot illustrating the log₂ fold changes in expression of 25 BCL11A target genes in BCL11A^E-disrupted erythroblasts generated in vivo (x-axis) vs in vitro (y-axis). Colored sections represent thresholds for twofold change (red) and 1.5-fold change (yellow). (D) GSEA of altered gene expression in BCL11A^E-disrupted vs control erythroblasts generated in vivo. Gene sets represented on the left and right halves of the graph are enriched in downregulated and upregulated genes, respectively. (E) GSEA comparing altered gene expression pathways in BCL11A^E-disrupted erythroblasts generated in vivo and in vitro. BasoE, basophilic erythroblast; log FC, log fold change.