Figure 3

Effect of Cl-amidine and TSA treatment on histone H3 and H4 acetylation and methylation. A. PN zygotes were cultured for ~68 hours in KSOM medium supplemented with either 250 μM of Cl-amidine or 100 nM of TSA. Two or four-cell embryos were fixed and immunostained with antibodies to hyperacetylated H4 (red) and acetyl H3K9 (green). DAPI was utilized to detect DNA (blue). DIC, differential interference contrast. B. Histograms documenting the fluorescent intensity of the hyperacetylated H4 and acetyl H3K9 signals shown in A. C. Same as (A) except antibodies against acetyl H3K18 (red) and di-methyl H3K9 (green) were used. D. Histograms documenting the fluorescent intensity shown in B experiments. E. Same as (A) except antibodies against acetyl H4K5 (red) were used. F. Histogram documenting the fluorescent intensity in experiment shown in E. Data are presented as mean + SEM. *P < 0.05, **P < 0.005, ***P < 0.001 (two-tailed paired Student's t-test).