Fig. 2

Introducing insA elevates γ-globin expression in CD34⁺ HSPCs. (a-b) Quantitative measurement of Gγ/Aγ ratio by quantitative real-time PCR (a) and HbF production by western blot (b) in control (Ctrl) and insA CD34⁺ HSPCs. #1–4 indicated the replicates. (c) Flow cytometry was performed to detect the HbF cells in control and insA CD34⁺ HSPCs. (d) Statistics result of the HbF cells in control and insA CD34⁺ HSPCs. (e-f) cell proliferation (e) and cell viability (f) analysis in control and edited CD34⁺ HSPCs. Ctrl indicated the control cells edited in the AAVS1 locus. (g) Quantitative measurement of HBG mRNA expression by quantitative real-time PCR in control and insA clone CD34⁺ HSPCs. Data from ≥2 independent experiments are presented as mean ± SD (*p < 0.05, ***p < 0.001, ****p < 0.0001)