Fig. 1: Effects of DNMT1 on proliferation of erythroid progenitors.

A Heatmap showing mRNA levels of DNMTs as assessed by RNA-seq in erythroblasts. B qRT-PCR results showing DNMT1 mRNA expression levels in erythroblasts cultured for 4, 7, 9,11,13,15, and 17 days. GAPDH was used as internal reference. C Representative western blots showing DNMT1 protein levels in erythroblasts cultured for 4,7,9,11,13,15 and 17 days. D Quantitative analysis of DNMT1 protein levels from 3 independent experiments. E qRT-PCR results showing DNMT1 mRNA expression levels in luciferase-shRNA or DNMT1-shRNAs transduced erythroblasts cultured for 6 days. GAPDH was used as internal reference. F Representative western blot showing DNMT1 protein levels in luciferase-shRNA or DNMT1-shRNAs transduced erythroblasts cultured for 6 days. G Quantitative analysis of DNMT1 protein levels from 3 independent experiments. H Growth curves of Luciferase and DNMT1-knockdown and DNMT1 inhibitor erythroblasts at first 6 days. I Representative flow cytometry profiles of cell cycle as assessed by Edu and 7AAD staining of cells cultured for 6 days. J Quantitative analysis of cell cycle. K Representative flow cytometry profiles of apoptosis as assessed by dual staining of Annexin V and 7AAD of cells cultured for day 6. L Quantitative analysis of apoptosis from 3 independent experiments. M Colony-forming ability of cells cultured for day 6, which contain mixed populations of cells that include BFU-E, CFU-E, and proerythroblasts. N Colony number of Control (luciferase-shRNA and N-DMSO) and DNMT1-deficient (DNMT1-shRNAs and DC_517 1 µM) on day 6 of culture. * p < 0.05, ** p < 0.01, *** p < 0.001.