Fig. 4

circRIP2 sponges miR-1305 to elevate Tgf-β2 in bladder cancer cell. a, b FISH and nuclear-plasma extraction assay were performed to detect the location of circRIP2. Cytoplasm located 18SRNA was taken as positive control, scale bar: 2.5 μm; c. Heat map and RNA-sequencing analyze mRNAs expression that were differently expressed between circRIP2 over-expressed and vector in U3 bladder cancer cells; Each group contains 3 samples; Each column corresponds to the expression profile of a sample, and each row represents a mRNA (log fold change (FC) ≥1.5 and P < 0.05); d. Western blot re-verified the up-regulation of Tgf-β2 caused by circRIP2 over-expression; e, f. RNA pull-down assay was used to evaluate miRNAs that could bind with circRIP2 in 2 bladder cancer cells, 5637 and U3; g. Pull-down assay for biotin labeled miRNA was used to evaluate binding properties between miR-1305 and circRIP2 in 2 bladder cancer cells, 5637 and U3; h. Dual luciferase reporter assay was used to prove the binding properties between circRIP2 and miR-1305; i. Western blot showed a significant interfering effect on Tgf-β2 expression by miR-1305; j. Dual luciferase reporter assay showed the binding property between miR-1305 and Tgf-β2;