Erythroid differentiation of CD34⁺cells. (A) CD34⁺-enriched cells were induced to erythroid differentiation in the absence or presence of DMK, ALA, nucleosides, or AOA. (B) Cells were exposed to DMK, ALA, nucleosides, or DES in combination with AOA as indicated. After 6 days of differentiation and treatment in StemSpan media with erythroid expansion supplement, cells were collected and differentiation was assessed by flow cytometry detection of cell surface markers CD235a and CD71. Labels for each treatment type are shown at the top of the figures. For each treatment type, from top to bottom, graphs illustrate flow cytometric pseudocolor plots of cells stained using CD235a (x-axis) and CD71 (y-axis) antibodies, histograms of these cells stained for CD71, and histograms of these cells stained for CD235a. (C) Results from CD34⁺ cells that were grown for 6 days in StemSpan media with erythroid expansion supplement before the removal of defined growth factors/cytokines other than EPO as described in the text. Cellular heme levels are shown for cultures after 3 and 6 days in the presence of EPO with indicated compound. UN, untreated. The bar graph presents combined biological and technical replicates (n = 4-6), mean ± standard deviation.