Fig. 3

The Expression and Activity of HDAC2 at the Initial Stage of Ovulation. A RT-qPCR results demonstrate the mRNA expression levels of HDACs (Hdac1-10) in ovaries at different timepoints. B The protein levels of HDAC1, HDAC2, pHDAC2, and Histone H3 were determined by Western blotting in ovaries at different timepoints. P0, P24: 0 h and 24 h after PMSG treatment. H0–H8: 0, 1, 2, 4, and 8 h after hCG treatment followed by PMSG treatment for 48 h. C, D Representative immunofluorescence for HDAC2 (red in C) and pHDAC2 (red in D) in large antral follicles at different timepoints after PMSG and hCG treatment. Scale Bar = 100 μm. H0, H1, and H4: 0 h, 1 h, and 4 h after hCG treatment followed by PMSG treatment for 48 h. White dotted circles indicate cumulus cells and oocyte complex (COC). E, F Quantities of HDAC2 and pHDAC2 signal intensity in the COC in panels C and D. Data are presented as the mean ± SD (two-tailed unpaired t-test). G The levels of HDAC2 activity in GCs at different timepoints were determined using an in vitro assay kit. P24: 24 h after PMSG treatment. H0–H4: 0, 1, 2, and 4 h after hCG treatment, followed by PMSG treatment for 48 h. All data are presented as the mean ± SD. Two-tailed unpaired t-tests were performed for experiments depicted in panels E–F. One-way ANOVA was performed, followed by Turkey’s multiple comparisons test for experiments depicted in panel G. NS, no significance, *P < 0.05, **P < 0.01, ***P < 0.001