Fig. 4: TRAF6 and IRAK1 are the miR-146a-5p target genes in GBM.

A Venn diagram (left panel) showed the intersection of predicted binding genes of miR-146a-5p obtained from 6 online public databases. Nine genes (middle panel) were predicted in all six databases. Association analysis of the proteins encoded by these 9 genes performed with STRING (right panel) showed a strong connection between TRAF6 and IRAK1. B Schematic representation of the TRAF6 3′UTR and IRAK1 3′UTR. MT sequences were generated at the 3′UTR binding sites of miR-146a-5p according to the prediction. C Relative luciferase activity was determined in U87MG cells that were cotransfected with the miR-146a-5p mimic or mimic NC and the WT-3′UTR or MT-3′UTR of TRAF6 or IRAK1. D Relative luciferase activity was determined in U87MG cells transfected with the WT-3′UTR or MT-3′UTR of TRAF6 or IRAK1 for 24 h and then cultured with M1-EVs or mock. E Western blotting measured the expression levels of TRAF6 and IRAK1 in U87MG and A172 cells cocultured with M1-EVs or M2-EVs. F Expression of TRAF6 and IRAK1 in U87MG and A172 cells transfected with the miR-146a-5p mimic, miR-146a-5p inhibitor, mimic NC, or inhibitor NC. G The correlation of TRAF6 and IRAK1 mRNA expression in patients with LGG and GBM in TCGA was estimated, and the mRNA expression level of TRAF6 was positively correlated with that of IRAK1 in GBM and LGG. H The DFS and OS of glioma patients with different TRAF6/IRAK1 expressions were analyzed in TCGA database. Both DFS and OS of LGG patients with higher TRAF6/IRAK1 expression levels were shorter than those with lower expression levels. Both DFS and OS of GBM patients did not obtain significant benefit.