FIG 2.

Setd8 knockdown in ESREs is not associated with abnormal proliferation, decreased viability, or accumulation of DNA damage. (A) During maturation, cell counts are similar in control and knockdown cultures. Data represent the means and SEM from a minimum of three independent experiments. (B) Trypan blue staining demonstrates similar viability in control and knockdown cultures. Data represent the means and SEM from a minimum of three independent experiments. (C) Image stream analyses using an antibody specific to γ-H2Ax did not detect significant numbers of γ-H2AX foci in knockdown or control cultures. A UV-treated sample is included as a positive control. (D) Setd8 knockdown is not associated with altered cell cycle progression in erythroid cells. Flow-cytometric analyses utilizing an antibody specific for Ki-67 demonstrate that the scramble control and knockdown cultures have similar proportions of cells in each phase of the cell cycle. The left panel is an example of the gating strategy. The bar graph demonstrates the percentage of cells in each phase of the cell cycle for the Setd8 knockdown and control cultures. The means and SEM from three independent experiments are shown.