Fig. 5

HDAC2-mediated Deacetylation is Essential for Transcription Switching and Successful Ovulation. A A diagram depicting the experimental design. FK228, the inhibitor of HDAC1/2, was pretreated for 4 h before hCG injection to induce ovulation. Black arrows indicate the time points when the ovaries were collected. H0–14: 0 h, 1 h, 4 h, 8 h, and 14 h after hCG treatment. B Western blotting of the main acetylated histones at different timepoints post-hCG in the control and FK228 groups. C Immunofluorescence of H3K27Ac at different timepoints (0, 1, 4, and 8 h) after hCG treatment in the control and FK228 groups. Scale bar = 100 μm. D The ovary morphology with hematoxylin and eosin staining from the control and FK228 groups at 14 h post-hCG. E The average oocyte numbers ovulated per mouse. The results are presented as the mean ± SD. ***P < 0.001, calculated by t-test. F ChIP-qPCR analysis of H3K27Ac on Ereg, Star, and Sult1e1 at 0 h and 4 h post-hCG pre-treated with or without FK228. G The relative mRNA levels by RT-qPCR of Ereg, Star, and Sult1e1 at 0, 4, and 8 h post-hCG in the control and FK228 groups. The fold change was represented by setting the relative level of 0 h in the control group as 1. The results (panels F and G) are presented as the mean ± SD. *P < 0.05, **P < 0.01, as calculated by one-way ANOVA, followed by Turkey’s multiple comparisons test on log-transformed values