Fig. 5From: Epigenetically silenced lncRNA SNAI3-AS1 promotes ferroptosis in glioma via perturbing the m6A-dependent recognition of Nrf2 mRNA mediated by SND1SNAI3-AS1 binds to SND1 protein. A RNA FISH analysis of SNAI3-AS1 localization in U87MG and A172 cells. 18S and U6 were used as positive controls. B After nucleocytoplasmic separation assay, the expression level of SNAI3-AS1 was determined via RT-qPCR. GAPDH and U6 were applied as positive controls. C Silver staining was used to identify the SNAI3-AS1-binding proteins pulled down by synthesized biotin-labeled SNAI3-AS1 probe. D Validation of the interaction between SNAI3-AS1 and SND1 protein through western blotting. E RIP assays were performed using anti-SND1 and IgG antibodies. The enrichments of SNAI3-AS1 by SND1 or IgG were detected via RT-qPCR. F The colocalization between SNAI3-AS1 and SND1 was determined by FISH combined with IF staining. G The predicted secondary structure of SNAI3-AS1. H Deletion mapping of the SND1-binding domain in SNAI3-AS1. Top, diagrams of full-length SNAI3-AS1 and the deletion fragments. Middle, the in vitro–transcribed full-length SNAI3-AS1 and deletion fragments with correct sizes were visualized by agarose gel image in U87MG cells. Bottom, immunoblot analysis for SND1 in the protein samples pulled down by different biotinylated SNAI3-AS1 truncations in U87MG cells. I The diagrams of Flag-tagged full-length or truncation plasmids with various assembled domains of SND1 protein. J Full-length or truncations of recombinant SND1 protein with correct sizes were validated by western blotting using anti-Flag in U87MG cells. K RT-qPCR detected the relative enrichment levels of SNAI3-AS1 in full-length or truncation SND1 RIP assays using anti-Flag and anti-IgG in U87MG cells. **P < 0.01, ***P < 0.001, and n.s., not significantBack to article page