Fig. 6

MAPK1 and E2F7 are direct targets of miR-129-5p and is suppressed by SNHG12 detection. a Schematic drawing of the screening procedure of candidate target genes. b After transfected with miR-NC or miR-129-5p in Rec GBM and N3T3rd cells, the expression level of 8 potential targets for miR-129-5p was analyzed using real-time PCR. c The luciferase reporter plasmid carrying wild type (WT) or mutant (MUT) MAPK1 or E2F7 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. d RIP experiments were performed using the Ago2 antibody, and specific primers were used to detect the enrichment of MAPK1 and E2F7. e RIP assay of the enrichment of Ago2 on SNHG12, MAPK1 and E2F7 transcripts relative to IgG in Pri GBM and N3S cells transfected with pcDNA-ctrl or pcDNA-SNHG12. f RIP assay of the enrichment of Ago2 on SNHG12, MAPK1 and E2F7 transcripts relative to IgG in Rec GBM and N3T3rd cells transfected sh-ctrl or sh-SNHG12. g Relative protein levels of E2F7 and MAPK1 in Rec GBM and N3T3rd cells transfected with control miRNA, miR-129-5p-inhibitor or miR-129-5p mimics. h MAPK1 and E2F7 protein level in Rec GBM and N3T3rd cells following knockdown of SNHG12. i and j Luciferase activity of reporters which contained wild-type or mt MAPK1 or E2F7 3’UTR with indicated treatment in Rec GBM cells (i) and Pri GBM cells (j) k Relative protein levels of E2F7 and MAPK1 in Rec GBM and N3T3rd cells following knockdown of SNHG12 and/or inhibition of miR-129-5p. l Western blot analysis of E2F7 and MAPK1 in Pri GBM cells and N3S cells transfected with pcDNA-Ctrl, pcDNA-SNHG12, or pcDNA-SNHG12mt along with miR-129-5p mimics. Data are presented as the mean ± SEM from three independent experiments. Significant results were presented as NS non-significant, **P<0.01, ***P<0.001