Deletion and pharmacological inhibition of PTEN induces γ-globin. (A) HbF induction in 2 independent PTEN-deficient clonal HuDEP2 cell lines (PTEN^−/− #35 and #54) compared with control cells (nontargeting-4), as demonstrated by HbF staining and flow cytometry (n = 3). (B) Analysis of γ-globin and β-globin mRNA levels in PTEN-deleted clonal cells (n = 3). (C-F) CD34⁺ cells isolated from healthy donors were differentiated into erythroid cells and treated with LSD1 (GSK690), EZH (Taze), or PTEN inhibitors (bpV), showing induction of HbF with all 3 compounds, as demonstrated by HbF staining and flow cytometry (C-D; n = 5) and mRNA analysis (E-F; n = 4). (G-H) CD34⁺ cells isolated from β-thalassemia patients were differentiated into erythroid cells and treated with EZH (Taze, n = 3) or PTEN (bpV, n = 6) inhibitors. Both compounds result in HbF induction by high-performance liquid chromatography analysis. Data are shown are means ± standard deviation (A-B,E-F: **P < .01; ***P < .001; n.s., not significant; unpaired Student t test; D,H: *P < .05; **P < .01; paired Student t test).