Figure 2.

Lentiviral knockdown of MBD2 in HUDEP-2 cells results in progressively increased γ-globin gene expression. (A) Schema of HUDEP-2 cell expansion for four, seven, or ten days after MBD2 shRNA lentiviral transduction, prior to a 3-day differentiation period. mRNA and protein were harvested after the expansion and differentiation period for all samples. (B) Western blot showing qualitative degree of MBD2 protein knockdown at the day-4 time point. (C) Time-course plot of γ-globin mRNA as a percentage of total globin mRNA by qRT-PCR at the indicated time points. (D) Relative MBD2 mRNA expression normalized by cyclophilin A compared to the shSCR sample by quantitative real-time polymerase chain reaction at four, seven, and ten-day time points. Error bars represent ± Standard Deviation of three biological repeats. *P<0.05; **P<0.01; n.s.: P>0.05 between sample and scramble. Statistical testing was performed using the Student’s t-test.