Fig. 5

SNHG12 act as a sponge for miR-129-5p in the cytoplasm. a FISH analysis indicated subcellular location of SNHG12 in Rec GBM and N3T3rd cells (green). Nuclei were stained by DAPI (blue). b Relative SNHG12 expression levels in nuclear and cytosolic fractions of Rec GBM and N3T3rd cells. U6 was used as nuclear controls. β-actin was used as cytosolic controls. c RIP experiments were performed using the Ago2 antibody, and specific primers were used to detect the enrichment of SNHG12 and miR-129-5p in Rec GBM and N3T3rd cells. d Schematic drawing of the screening procedure of candidate miRNAs. e The luciferase reporter plasmids carrying SNHG12 was co-transfected into HEK293T cells with 5 miRNA-coding plasmids. f Up: Schematic representation of the miR-129-5p binding sites in SNHG12 and the site mutagenesis. Down: The luciferase reporter plasmid carrying wild type (WT) or mutant (MUT) SNHG12 was co-transfected into Rec GBM and N3T3rd cells with miR-129-5p in parallel with an empty vector. Relative luciferase activity in Rec GBM and N3T3rd cells were determined. g Colony formation ability of Rec GBM and N3T3rd cells transfected with SNHG12 plasmid, miR-129-5p mimics + SNHG12 plasmid or miR-129-5p inhibitor + SNHG12 plasmid after 200 μM TMZ treatment for 48 h. h Immunofluorescent staining of cleaved caspase-3 in Rec GBM cells transfected with SNHG12 plasmid, miR-129-5p mimics + SNHG12 plasmid or miR-129-5p inhibitor + SNHG12 plasmid after 200 μM TMZ treatment for 48 h. Scale bar = 50 μm. Data are presented as the mean ± SEM from three independent experiments. Significant results were presented as NS non-significant, *P<0.05, **P<0.01