Fig. 4
From: ISG15 suppresses ovulation and female fertility by ISGylating ADAMTS1
ISG15 negatively regulates ovulation-related genes expression in an ISGylation-dependent way in mGCs. A, B Quantitative RT-PCR (A) analysis and western blot (B) analysis of ovulation-related genes including Adamts1, Ptgs2, Has2, Areg, Ereg in mGCs overexpressing Isg15 and Isg15aa. mGCs were transfected with pcDNA3.1, pcDNA3.1-Isg15 or pcDNA3.1-Isg15aa (ISG15 conjugation-defective mutant) for 48 h and cell lysates were collected for assays. C ISGylation level in mGCs overexpressing Isg15 and Isg15aa. mGCs were transfected with pcDNA3.1, pcDNA3.1-Isg15 or pcDNA3.1-Isg15aa for 48 h and cell lysates were collected for assays. D, E Quantitative RT-PCR (D) analysis and western blot (E) analysis of ovulation-related genes including Adamts1, Ptgs2, Has2, Areg, Ereg, Isg15 in mGCs overexpressing E1 activating enzyme Uba7. F, G Quantitative RT-PCR (F) analysis and western blot (G) analysis of ovulation-related genes including Adamts1, Ptgs2, Has2, Areg, Ereg, Isg15 in mGCs overexpressing E2 conjugating enzyme Ube2l6. H, I Quantitative RT-PCR (H) analysis and western blot (I) analysis of ovulation-related genes including Adamts1, Ptgs2, Has2, Areg, Ereg, Isg15 in mGCs overexpressing E3 ligating enzyme Herc6. J, K Quantitative RT-PCR (J) analysis and western blot (K) analysis of ovulation-related genes including Adamts1, Ptgs2, Has2, Areg, Ereg, Isg15 in mGCs overexpressing E3 ligating enzyme Trim25. L, M Quantitative RT-PCR (L) analysis and western blot (M) analysis of ovulation-related genes including Adamts1, Ptgs2, Has2, Areg, Ereg, Isg15 in mGCs overexpressing ISGylation-specific deconjugating enzyme Usp18. All data are mean ± SD; *P < 0.05, **P < 0.01