The HBB:c-78A>G mutation leads to reactivation of γ-globin in HUDEP-2 cells and CD34⁺ cells. (A) Quantitative measurement of HBG mRNA expression with the HUDEP-2 cells and HBB:c.-78A>G single clone by qPCR. (B) WB in HUDEP-2 WT cells with the HBB:c.-78A>G single clone. (C) Hb F production by flow cytometry from HUDEP-2 WT cells and HBB:c.-78A>G single clone. (D) Hb F–positive cells analyzed by flow cytometry are represented as upper histograms referring to edited 3 healthy donors CD34⁺ cells. The lower histograms refer to edited 3 patients with β⁰/β⁰ CD34⁺ cells on day 16 of differentiation (n = 3). (E) Flow cytometry analysis of Hb F–positive cells in erythroblasts derived from edited 3 healthy donors and 3 patients with β-thalassemia CD34⁺ cells on day 16 of differentiation (n = 3). Asterisks indicate levels of statistical significance. ∗P < .05. FITC-A, fluorescein isothiocyanate-area; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; mRNA, messenger RNA; SSC-A, side scatter-area.