FIGURE 6.

PCAF knockdown inhibits ATRA-induced granulocytic differentiation in HL-60 cells. A and B, HL-60 cells expressing DsRed were sorted, and knockdown efficiency was analyzed by RT-qPCR (A) and immunoblotting (B). Three independent sequences of shRNAs (the same sequences as in Fig. 5) were used against PCAF. The RT-qPCR results were quantified as described in the legend to Fig. 5. Experiments were performed in triplicate, and the data are shown as the mean ± S.D. (error bars). For the immunoblotting, 4.2 μg of extract was loaded into each lane. C, PCAF knockdown HL-60 cells and control cells expressing non-targeting shRNA were cultured with 10 nm ATRA or ethanol (vehicle) for 72 h. FACS analysis for monitoring DsRed and CD11b expression was then performed. Infection efficiency was ∼20–40% based on the proportion of cells expressing DsRed.