Fig. 4

LINC00612 served as a molecular sponge for miR-590. a Biotinylated miRNA were transfected into T24 cells. RT-qPCR was performed to quantify the RNA levels of LINC00612 and GAPDH. A scatter plot showing the relative ratios of the input of IP. *P < 0.05. b A schematic diagram showing the putative miR-590 binding sites with the LINC00612. The sequences of wild-type LINC00612 and mutant LINC00612 are also listed. Luciferase reporter gene assays were performed to measure the luciferase activity in T24 cells. *P < 0.01. c Biotinylated miR-590 or its mutant (miR-590-mut) was transfected into T24 cells. RT-qPCR was performed to quantify the RNA levels of LINC00612 and GAPDH. A scatter plot showed the relative ratios of the input of IP. *P < 0.01. d Anti-AGO2 RIP assays were used in T24 cells to determine LINC00612 and miR-590 RNA enrichment in IP complexes. Anti-IgG was used as a control. *P < 0.01. e Relative expression of miR-590 in BC cells that were transfected with the Lv-LINC00612 vector/Lv-NC vector and shLINC00612 vector/shNC vector were measured via RT-qPCR. f RT-qPCR was performed to measure the relative expression of pre-miR590 and mature-miR590 in BC cell lines (5637, UMUC3, and T24) and human bladder epithelium immortalized cells (SV-HUC-1). **P < 0.01. N = 3 independent experiments