Additional file 1 of Ovulatory signal-triggered chromatin remodeling in ovarian granulosa cells by HDAC2 phosphorylation activation-mediated histone deacetylation
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- Jin, Jiamin; Ren, Peipei; Li, Xiang; Zhang, Yinyi; Yang, Weijie; Ma, Yerong; Lai, Mengru; Yu, Chao; Zhang, Songying; Zhang, Yin-Li
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Additional file 1. Figure S1. The Dynamics of H3K27Ac Levels during Follicular Growth and Ovulation. Mice were treated with pregnant mare serum gonadotrophin (PMSG) to stimulate follicle growth or human chorionic gonadotropin (hCG) to trigger ovulation. Ovaries were collected at the indicated timepoints for immunofluorescence. The representative images showing the H3K27Ac (red) levels with DAPI (blue) co-stained for visualization of nucleus (n=3). Scale bar=100um. Figure S2. The H3K27Ac ChIP-seq Analysis for H3K27Ac-gain Genes after Ovulation Signal Induction. (A) Pie charts represent the ratio of stable H3K27Ac-enriched peaks, H3K27-loss peaks and H3K27-gain peakss (de novo H3K27Ac-deposited peaks and H3K27Ac-increased peaks). (B) Genome browser snapshot shows Prss56 is one of de novo H3K27Ac-gain genes after hCG induction. (C) Gene Ontology (GO) analysis shows the biological process (BP) of de novo H3K27Ac-gain genes after hCG induction. (D) Bar charts representing the top enriched KEGG pathway of the ovulatory specific genes 4 h post hCG. (E) The HOMER known and de novo motif analysis for the H3K27Ac-gain peaks to predict transcriptional factors. Figure S3. Semiquantitative analysis of protein levels in Fig. 3. The western blot band intensities of Fig. 3B were measured with ImageJ software. HDAC1 and p-HDAC2 protein levels were normalized to Histone H3 levels. Data were expressed as mean±SD. P value was determined by two-way ANOVA followed by Tukey’s post-test. ** P