Research Paper Volume 12, Issue 3 pp 3010—3024

microRNA-1203 targets and silences cyclophilin D to protect human endometrial cells from oxygen and glucose deprivation-re-oxygenation

Figure 1. miR-1203 targets and silences CypD in human endometrial cells. The wild-type (WT) microRNA-1203 (miR-1203) targets CypD 3’-UTR (3’-untranslated region) at position 806-813 (A). T-HESC endometrial cells were infected with pre-miR-1203-encoding lentivirus (“lv-pre-miR-1203”), following puromycin selection three stable cell lines were established: “sL1/sL2/sL3”. Control T-HESC cells were infected with microRNA control lentivirus (“lv-miRC”); Expression of mature miR-1203 and CypD mRNA was tested by qPCR assays (B and D); The relative CypD 3’-UTR luciferase reporter activity was examined (C), with CypD protein expression tested by Western blotting assays (E). T-HESC cells were transfected with 500 nM of control microRNA mimic (“miRC”), the wild-type (“WT-”) or the mutant (“Mut1/2”, see sequences in A) miR-1203 mimics for 48h, the relative CypD 3’-UTR luciferase reporter activity (F), CypD mRNA (G) and protein (H) levels were tested. The primary human endometrial cells (“Endometrial cells”, same for all Figures) were infected with lv-pre-miR-1203 or lv-miRC lentivirus for 48h, expression of mature miR-1203 (I), CypD mRNA (J) and protein (K) was shown. CypD protein expression was quantified and normalized to the loading control (E, H and K). “MW” stands for molecular weight (same for all Figures). “Vec” stands for the empty vector control (same for all Figures). Data were presented as mean ± SD (n=5). * P <0.05 vs. “Vec”/“miRC”/“lv-miRC” cells. Experiments in this figure were repeated three times with similar results obtained.