Figure 6.

Lentiviral shRNA knockdown (Kd) of MBD2 in CD34 progenitor-derived primary human erythroid cells results in high level γ/γ+β RNA expression and γ-globin protein without affecting erythroid differentiation. (A) Relative Kd of MBD2 mRNA’. (B) approximately 10-fold increase in γ/γ+β mRNA in MBD2 kd primary erythroid cells, across two different levels of differentiation, compared to scrambled shRNA controls. (C) Increase in γ-globin protein without change in β-globin protein as measured by western blot using anti γ-globin and anti β-globin antibody. (D) Flow cytometry analysis showing equivalent erythroid differentiation profiles of scramble control (sc) and MBD2 Kd CD34⁺ cells at day 7. Error bars represent ± Standard Deviation of three biological repeats. *P<0.05; **P< 0.01; n.s.: P>0.05. Statistical testing was performed using the Student’s t-test.