Fig. 4

HDAC2 is Essential for Histone Deacetylation, Ovulation-specific Genes’ Transcription, and Cumulus Expansion. A Western blotting results demonstrating the dynamics of pHDAC2 and the increased H3K27Ac level with the transfection of Hdca2 siRNAs in mouse primary GCs. Primary GCs were transfected with siRNAs against negative control (siNC) or Hdac2 (siHdac2) for 24 h, followed by the addition of 10 uM Forskolin (FSK) and 20 nM PMA to activate the ovulatory signal. B The representative images show the cumulus cell expansion capacity in the si-NC and si-Hdac2 groups. C The Hdac2 mRNA level was determined by RT-qPCR in cumulus cells derived from COCs of the si-NC and si-Hdac2 groups. The data are presented as the mean ± SD with a t-test on log-transformed values. ***P < 0.001. D Quantification of the indicated ovulatory specific genes expression in primary GCs by RT-qPCR following ovulatory signal in the si-NC and si-Hdac2 groups. E The RT-qPCR results for ovulatory specific genes’ mRNA levels in COCs from the si-NC and si-Hdac2 groups. The data are presented as the mean ± SD. One-way ANOVA followed by Turkey’s multiple comparisons test on log-transformed values. NS, no significance, *P < 0.05, ***P < 0.001, ANOVA, analysis of variance. P0, P24, H0: 0 h, 24 h, and 48 h after PMSG treatment. H1–H8: 1, 2, 4, and 8 h post-hCG treatment