Research Paper Volume 11, Issue 23 pp 10826—10838

Figure 4. circ5912 reverses Tgf-β2-induced EMT in bladder cancer. To identify the underlying mechanism of circ5912, (A, B) The mRNA sequence was analyzed for the downstream alteration after circ5912 overexpression; cells were treated with TGF-β2 in 10ng/ml for 24 hours to induce EMT. (C) the bladder cancer phenotype was observed under light microscope, scale bar: 2.5μm; (D) western blot analysis was performed to determine the levels of different endothelial and mesenchymal markers; (E) FISH and (F) nuclear-plasma extraction assays were performed to detect the location of circ5912, scale bar: 2.5μm. Cytoplasmic 18s RNA was used as a positive control; (G, H) RNA pulldown assay showed the binding properties of multiple miRNAs and circ5912.