Fig. 5

miR-941 fails to affect OGDR-induced cytotoxicity in Keap1/Nrf2-KO human endometrial cells. Stable T-HESC cells with the CRISPR/Cas9-Nrf2-KO construct (“Nrf2-KO”) (a-d) or the CRISPR/Cas9-Keap1-KO construct (“Keap1-KO”) (e-h) were further infected with the pre-microRNA-941 anti-sense lentivirus (“antagomiR-941”), the pre-microRNA-941 lentivirus (“lv-pre-miR-941”) or non-sense microRNA lentivirus (“lv-miRC”) for 48 h. These cells or the control cells with empty CRISPR/Cas9-KO construct (“Cas9-C”) were treated with OGD for 4 h, followed by re-oxygenation (“OGDR”) for 24 h, cell survival (a and e) and necrosis (b and f) were tested by CCK-8 and LDH release assays, respectively. Expression of mature miR-941 (c and g) and listed proteins (in total cell lysates, d and h) were shown. The lv-pre-miR-941-expressing T-HESC cells were further transfected with the UTR-depleted Keap1 construct (“+Keap1”) for 48 h, expression of listed genes was shown (i and j); Cells were subjected to the same OGDR stimulation for 24 h, cell necrosis (testing medium LDH release, k) and mature miR-941 expression (l) were tested similarly. Keap1 and Nrf2 protein expression was quantified and normalized to Tubulin (d, h and j). Data were presented as mean ± SD (n = 5), and results were normalized.. ^#P < 0.05 vs. OGDR treatment in “Cas9-C” cells (a-c, e-g). ^#P < 0.05 (i and k). Experiments in this figure were repeated three times with similar results obtained