Fig 1: Identification of FOS and its regulation of neurotrophins in placenta of polycystic ovary syndrome (PCOS). (a) Heatmaps of the global DNA methylation in the promoter and gene expression data for placentae from women with PCOS and controls. (n = 3 in each group) (b) Scatter plot of global DNA methylation levels in placentae in controls (x-axis) compared to women with PCOS (y-axis). Point density is shown as blue shading; Peacock blue and purple dots indicate significant down-regulated and up-regulated cytosine phosphate guanine (CpG) sites associated with a methylation difference > 0.20. (c) Volcano plot to inspect differentially expressed genes in placentae from women with PCOS and controls, with fold change as the abscissa and -log10 (P value) as the ordinate. Peacock blue and purple splashes represent genes that significantly up or down regulated respectively. Gray splashes mean genes without significantly different expression. FOS gene is marked as red. (d) Gene Ontology (GO) terms enriched in differentially expressed genes in placentae from women with PCOS. (e) Neural GO terms enriched in differentially expressed gene (top) and methylated DNA (bottom) in placentae from women with PCOS. (f) FOS gene mRNA and protein expression levels in human placentae, as assessed by real-time polymerase chain reaction (real-time PCR) (PCOS: n = 20; control: n = 32) and immunofluorescence with FOS (green), and nuclear DNA (blue) labeling (n = 5 in each group). All scale bars are 50 µm. (g) The summary data of methylation status at the promoter region of FOS gene in the placental villi using bisulfite genomic sequencing (BSP) (n = 3 in each group, significance was determined by Chi-square test). (h) In silico analysis of the genomic DNA 5 kb upstream and downstream of BDNF (pIV) and NGF (pI) transcription start site (TSS). Detection of FOS (i) protein expression levels, BDNF (j) and NGF (k) mRNA expression levels in overexpression group (n = 3) and control group (n = 3). Data were presented as mean ± SEM. *P < 0.05, unpaired two-tailed Student's t tests.
Fig 2: Brain derived neurotrophic factor (BDNF) and nerve growth factor (NGF) in placental villi. (a) Visualization of BDNF and NGF in placental villi of polycystic ovarian syndrome (PCOS) and control group by immunofluorescence with BDNF (green) and NGF (red). Nuclei were counterstained with 4',6-diamidino-2-phenylindole (DAPI) (blue fluorescence). The syncytiotrophoblasts layers are labeled with white arrows, and the results of protein quantification are shown by bar plots (n = 5 in each group). All scale bars are 50 µm. BDNF and NGF mRNA expression level were quantified by real-time polymerase chain reaction (real-time PCR). PCOS: n = 20; control: n = 32. Data were presented as mean ± SEM. *P < 0.05, unpaired two-tailed Student's t tests. (b) The methylation status at the promoter region of BDNF (blue) and NGF (purple) genes in the placental villi from women with PCOS and controls using bisulfite genomic sequencing (BSP) (n = 3 in each group, significance was determined by Chi-square test).
Supplier Page from CUSABIO Technology LLC for Human Nerve growth factor,NGF ELISA Kit