Fig. 7
SNAI3-AS1 perturbed the recognition of Nrf2 3’UTR by SND1 to exert ferroptosis-sensitizing activity. A RIP assays showed the enrichments of Nrf2 mRNA by SND1 in Nrf2 mRNA in U87MG cells with stable SNAI3-AS1 overexpression or A172 cells with stable SNAI3-AS1 knockdown. B After actinomycin D (5 μg/ml) treatment for 0, 2, 4, 6 h, RT-qPCR was used to analysis the Nrf2 mRNA stability in U87MG and A172 cells with indicated interventions. C Dual luciferase reporter plasmids containing Wild-type or mutant Nrf2 mRNA 3’UTR p were transfected into U87MG and A172 cells with indicated interventions, respectively. The relative luciferase activity was measured and normalized. D RT-qPCR detected the mRNA level of Nrf2 in U87MG and A172 cells with indicated interventions. E After DMSO or erastin (10 μM, 48 h) treatments, the protein levels of Nrf2 in U87MG and A172 cells with indicated interventions were determined by western blotting. F-I U87MG and A172 cells with indicated interventions were treated with erastin (10 μM) ± ferrostatin-1 (2 μM) for 48 h, cell viabilities were detected via CCK8 assays (F), intracellular MDA was determined by MDA assays (G), intracellular Fe2+ was measured by iron detection assays (H), lipid ROS accumulation was analyzed by flow cytometry with C11-BODIPY staining (I). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, and n.s., not significant