Fig. 4

SUV39H1 is a direct target of cancer cell-derived EV miR-744. a Venn diagram showing the target genes of miR-744 predicted by three online tools (miRSearch, TargetScan, and miRWalk). (B and C) Data obtained from the TCGA database analyzed for the expression of BIN1 (b) and SUV39H1 (c) in lung tumor and normal samples. The x-axis represents the sample type and the y-axis represents the gene expression value. d Putative miR-744 binding sites in the 3′UTR of SUV39H1 mRNA in the TargetScan website (http://www.targetscan.org/vert_71/). e Dual-luciferase reporter assay validating the interaction between miR-744 and SUV39H1 in HEK293T cells. f and g Expression of miR-744 (f) and SUV39H1 (g) determined by RT-qPCR in A549 and H460 cells treated with miR-744 mimic or miR-744 inhibitor. h Western blot analysis of SUV39H1 protein in A549 and H460 cells transfected with miR-744 mimic or miR-744 inhibitor. i and j Expression of miR-744 (i) and SUV39H1 (j) determined by RT-qPCR in A549 and H460 cells treated with BEAS-2B-, H1299-, and H522-derived EVs. k Western blot analysis of SUV39H1 protein in A549 and H460 cells treated with BEAS-2B-, H1299-, and H522-derived EVs. l Western blot analysis of SUV39H1 protein in A549 and H460 cells treated with EVs isolated from the BEAS-2B cells transfected with miR-744 inhibitor. (* p < 0.05, vs. BEAS-2B-derived EVs, or A549, H460 cells, or HEK293T cells transfected with mimic-NC; # p < 0.05, vs. A549 and H460 cells transfected with inhibitor-NC). Data are shown as mean ± standard deviation of three technical replicates. Data from two groups were compared using unpaired t-test and data from multiple groups were compared by one-way ANOVA with Tukey's post hoc tests