Figure 7.
Phosphorylation of H3S10 occurs mainly on hypoacetylaed H3 tails. (A) 293T cells were synchronized with nocodazole for 6 h to produce ~40% G2/M cells. The cells were acid-extracted, and 10 μg of extract was incubated with anti- H3ac antibody. The supernatant (Sup) and immunoprecipitate (IP) fractions were analyzed for phosphorylation and acetylation levels by Western blot. (B) Relative abundance of total unmodified H3K9, trimethylated H3K9 (H3K9me3), as well as acetylated H3K9 or H3K14 (H3K9ac or K3K14ac) peptides (open bars), with the percentage of each modified peptide in combination with H3S10 phosphorylation in the phosphorylated peptide population (black bars) isolated from nocodazole-synchronized mitotic HeLa cells as determined by mass spectrometry (Supplementary Fig. 8c).