Fig. 6: miR-596-3p suppresses brain metastasis by targeting IL-8.

A Conditioned medium from PC9-BrM cells that were transfected with miR-596-3p/miR-NC were conducted by cytokine array (Abcam) analysis. Fold change (FC) was examined by using Image J software. B PC9-BrM and H1915-BrM cells were transfected with miR-596-3p/miR-NC, and the expression of IL-8 was determined by qRT-PCR assay. C PC9-BrM cell was transfected with miR-596-3p, and the level of IL-8 was detected by immunofluorescence assay. D PC9-BrM cells was treated with miR-596-3p inhibitor, miR-inhibitor NC or miR inhibitor+YAP1, and the level of IL-8 was detected by qRT-PCR. E Analysis of the luciferase activity of 293 T cell co-transfected with miR-596-3p mimics/miR-NC and the luciferase reporter psi-CHECK2-YAP1-WT/MUT. F PC9 parental cells were cultured on the upper side of a transwell chamber by adding with various doses of recombinant IL-8, and the number of invaded cells was calculated after 24 h. G PC9 cells with increased IL-8 were added into the transwell insert, and the number of invaded cells was calculated after 24 h. H HBMEC cells were added with conditioned medium from PC9-BrM, 231BrM+miR-596-3p and recombinant IL-8, the transendothelial electrical resistance was detected by EVOM2 epithelial voltohmmeter.