Exploration and validation of genes with androgen receptor binding sites and regulating mitochondrial biogenesis in HTR-8/SVneo cells and villi. A, Target genes with androgen receptor binding sites and meanwhile regulating mitochondrial biogenesis were screened using the Gene Transcription Regulation Database (GTRD) and MitoCarta3.0 database. B, Quantitative analysis for target genes messenger RNA (mRNA) expression by real-time quantitative polymerase chain reaction in HTR-8/SVneo cells with or without dihydrotestosterone (DHT) stimulation. C, The protein level of mitochondrial transcription factor A (TFAM) was detected using Western blot in HTR-8/SVneo cells treated with DHT or not. D, Gray scale analysis of Fig. 4C. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as the loading control. E, Representative immunofluorescence images showing the expression pattern of TFAM (green) in trophoblast cells with or without β–human chorionic gonadotropin (red) expression from the control group and polycystic ovary syndrome (PCOS) patients with euploid miscarriage group. Nucleus were stained with DAPI (4′,6-diamidino-2-phenylindole) (blue). Scale bar: 50 μm. F, Quantitative analysis for TFAM mRNA in villi from the control group (n= 15) and PCOS patients with euploid miscarriage group (n = 8). G, The TFAM protein level was detected using Western blot in villi from the control group (n = 6) and PCOS patients with euploid miscarriage group (n = 7). H, Gray scale analysis of Fig. 4G. GAPDH was used as the loading control. *P less than .05; **P less than .01; ***P less than .001.