Fig 2: (A) Picture of pelleted K562 infected with an empty short hairpin RNA (shRNA, shCT) and two shRNAs against PHF19 (sh#1 and sh#2) 5 days after puromycin selection. (B) CD235a marker levels measured by flow cytometry in shCT-, sh# 1-, sh#2-infected cells 4 days after puromycin selection and 1 µM of Ara-C treatment in uninfected cells as a positive control. Left, representative plot of a single experiment. Right, bar plot of the mean of median cytometry values (shCt and sh#1, n = 4; sh#1 and 1 µM of Ara-C, n = 3) + SEM. (C) CD61 marker levels measured by flow cytometry in shCT-, sh# 1-, and sh#2-infected cells 4 days after puromycin selection and treated with 1 nM phorbol myristate acetate (PMA) for 24 h. Left, representative plot of a single experiment. Right, bar plot of the mean of median cytometry values (n = 3) + SEM. (D) CD235a marker levels measured by flow cytometry at 4 days in shCT-, sh# 1-, and sh#2-infected cells after puromycin selection and treated with 10 nM of Ara-C for 72 h, with 1 µM of Ara-C treatment in uninfected cells as a positive control. Left, representative plot of a single experiment. Right, bar plot of the mean of median cytometry values (shCt, 10 nM Ara-C; sh#1, 10 nM Ara-C, n = 24; sh#1, 10 nM Ara-C, and 1 µM of Ara-C, n = 3) + SEM. *?p<0.05;p**<0.01.

Fig 3: iPSCs Derived from Different Donors Show Variable Erythroid Lineage Differentiation Propensity(A) Representative micrographs showing the hematopoietic cells differentiated from iPSCs with reduced hemoglobinized erythroid cells in iPSCs derived from T42 (left panel). Scale bar, 100 µm.(B) Representative FACS plots of differentiated genetically matched F- and B-iPSC lines (n = 4) showing the percentage of erythroblasts (GPA+ CD71+ gated from CD45- CD33-).(C) Scatter graphs presenting percentage of erythroblasts generated from the genetically matched F- and B-iPSC lines. Each dot represents an independent experiment.(D) Percentage of erythroid colonies out of the total colony number generated from iPSC lines. Values are mean ± SEM. Statistics performed by Student's t test, in three or four biological replicates.(E) Gene set enrichment plot and heatmap showing the genes constituting the core enrichment of overlap between multi-potent progenitors sorted from the bone marrow of DBA patients and iPSC lines derived from T42 cells. T42 iPSC lines show decreased expression levels of genes that are downregulated in Diamond-Blackfan anemia patients.