Reduction of hemogen suppresses expression of genes associated with erythrocyte maturation and hemoglobinization in human CD34⁺ cells and HUDEP2 cells. (A) Schematic diagram of short hairpin RNA (shRNA) KD of hemogen in CD34⁺ human umbilical cord erythroid progenitor cells. (B) Volcano plot showing differential gene expression between WT and hemogen-KD CD34⁺ cells at differentiation day 10. Downregulated and upregulated genes are defined by an adjusted P value <.1 and log₂ fold change less than −0.6 or >0.6, respectively. Light blue dots highlight erythroid genes that were significantly downregulated upon hemogen KD. (C) GO terms enriched among genes that were significantly up- or downregulated upon hemogen KD in CD34⁺ cells at differentiation day 10. (D) Expression of erythroid-related and globin genes is shown as log₂ fold change in hemogen-KD cells relative to control CD34⁺ cells during differentiation days 10 to 14. (E) Schematic diagram of shRNA-mediated KD of hemogen in human HUDEP2 cells. (F) Venn diagram of overlapped genes that were downregulated upon hemogen KD at differentiation days 3, 5, and 7. (G) GO analysis of overlapped genes from (F). (H) Examples of genes that were downregulated by hemogen KD in differentiated HUDEP2 cells from (F).