Figure 5. SLC2A1 is the direct target of miR-140-5p. (A) Venn diagram containing genes that were predicted to be the targets of miR-140-5p, and the KEGG pathway analysis of the 45 putative genes which were in the intersection. Red represented reported targets of miR-140-5p. (B) The expression levels of LAMC1, SLC2A1 and FGF9 in MDA-MB-231 cells transfected with miR-140-5p mimic compared with NC mimic for 48h. (C) The transfection of miR-140-5p in MDA-MB-231 cells decreased the SLC2A1 protein levels, as shown by western blot. (D) Left, luciferase-SLC2A1 3′-UTR constructs. 3 putative miR-140-5p binding sequences existed in the 3′-UTR of SLC2A1 mRNA, one was conservative, and the other two were poorly-conservative. miR-140-5p seed mutated sequences were generated in the binding site. Right, luciferase reporter assay in HEK293 cells transfected with NC, miR-140-5p or miR-140-5p-mut 3′-UTR. Firefly luciferase served as an internal control. (E) Expression patterns of miR-140-5p with SLC2A1 exhibited a negative correlation. (F) The expression level of miR-140-5p was decreased in BRCA compared with normal tissues. This difference was also reflected in ACC, but there is no significant disparity in THCA and RRAD. The p-values were calculated using standard Student t-tests. Error bars represent mean±SEM of three individual experiments. * P ≤ 0.05.