EKLF-driven PIT1 expression is mandatory for erythroid maturation in vitro. (A) EKLF and PIT1 mRNA quantification during G1E-ER-Gata1 maturation. Means ± SEM of at least 3 experiments are presented. Significant differences from t = 0 are indicated. (B) Western blot of PIT1 and EKLF during G1E-ER-Gata1 maturation. (C) PIT1 and EKLF mRNA levels on murine G1E-ER-Gata1 erythroid cells transduced with lentivirus containing a nontargeting shRNA or shRNAs directed against PIT1 or EKLF. (D) Western blot of PIT1 and EKLF on murine G1E-ER-Gata1–transduced cells. (E-F) PIT1 (E) and EKLF (F) mRNA relative expression in G1E-ER-Gata1–transduced cells 24 hours (EKLF) and 48 hours (PIT1) after maturation induction. Results are normalized to scramble at t = 0 and are expressed as the percentage induction compared with scramble at t = 24 or 48 hours. (G,I) Dematin mRNA (G) and β-globin (I) expression in G1E-ER-Gata1–transduced cells 48 hours after maturation induction. Results are normalized to scramble at t = 0 and are expressed as the percentage induction compared with scramble at t = 48 hours. (H) Percentage of benzidine-positive G1E-ER-Gata1–transduced cells 48 hours after maturation induction. (J) Percentage of benzidine-positive cells 48 hours after maturation induction of EKLF-depleted G1E-ER-Gata1 cells transduced with PIT1 coding lentivirus (PIT1-GFP) or with a control lentivirus (IRES-GFP). (K-L) β-Globin (K) and dematin (L) mRNA expression in G1E-ER-Gata1 cells 48 hours after maturation induction. Results are normalized with control at t = 0 and are expressed as the percentage induction compared with PIT1-GFP at t = 48 hours. Means ± SEM of 5 independent experiments are presented. Significant differences from scramble at t = 48 hours are indicated. *P < .05; **P < .01; ***P < .001.