Figure 6.

Macrohistone mH2A2 is regulated by Plcγ1 and is involved in erythroid differentiation. (a) Quantitative RT-PCR of H2afy2 mRNA in I/11 cells after infection with either mH2A2 shRNA or control shRNA. Each experiment (n=3) was done in triplicate and the error bars represent mean ±S.D. (b) Differentiation of I/11 cells stably infected with either mH2A2 shRNAs or control shRNA was measured for 96 h (day 4) in response to Epo. A representative FACS blot (left panel) and percentage of immature (TER119⁺CD44^high) cells after 96 h (day 4; right panel) is shown. The error bars represent mean ±S.D. (n=4). (c) Representative cytospin analysis followed by Giemsa staining of mH2A2-deficient and control cells induced to differentiate in response to Epo. The scale bar represents 5 μm. (d) Survival of I/11 cells stably infected with either mH2A2 shRNA or control shRNA using Annexin V/SYTOX Blue staining. The error bars represent mean ±S.D. (n=3)