Fig. 2

Two-step inhibition of OGT and OGA induces erythroid differentiation and enucleation. A Schematic diagram of the treatment strategy developed based on the cellular O-GlcNAc level during erythroid differentiation from CD34⁺ HSPCs. Treatment was either OGAinh/OGTinh or OGTinh/OGAinh, replacing every 2 days and switching on day 11, for a total of 15 days—alloxan (ALX) was used at 1 mM, thiamet G (TG) was at 10 μM, and OSMI-1 was at 5 μM. B Percentages of differentiated erythroblasts in stages I–V as analyzed by flow cytometry using FSC versus CD71, derived from HSPCs of donors #7, #14, and #23 on day 15 of culture. C Representative morphology of treated cells on Wright-stained cytospin slides on day 15 of culture. Percentage of enucleated cells from at least 300 cells is indicated on the micrograph. Scale bar = 20 μm. D qPCR of genes involved in erythroid development and function in the differentiated cells derived from CD34⁺ HSPCs of donors #7 and #23 on day 15 of culture. Data were normalized to GAPDH and represented in a heatmap