Effects of GATA-2 expression on epigenetic modification and expression of PU.1. (A-C) On day 5, TSA was added and GATA-2 expression was induced. (A) Surface expression patterns of Mac-1, GPV, and TER-119 at day 9. (B) May-Giemsa staining at day 9. (C) RT-PCR analysis of PU.1, Gata1, and Gata2 expression at days 6 to 8. Serially diluted cDNAs (1, 1:3, and 1:9) were used for PCR. (D) Schematic PU.1 genomic locus and the primers used for ChIP analyses. GATA-2 expression was induced at day 5 or 6 as indicated, and ChIP analyses were carried out 8 hours later. (E) ChIP analysis using anti-GATA-2 antibody. (F) ChIP analysis of ES cells and differentiated cells using antiacetylated histone H4 antibody. (G) Effects of TSA on the amount of acetylated histone H4. GATA-2 expression commenced on day 5 and ChIP analyses were carried out 8 hours later. (H) Suppression of PU.1 promoter activity by GATA-2. The data are shown as the mean ± SD of 6 samples.