Fig. 6

Phenotypes of Caki-1 and 786-O cells cotransfected with siRNAs for circPDHK1 and PPP1CA. A Colony formation assays, (B) CCK-8 assays and (C, D) EdU assays were performed to detect the cell viability and proliferation activity of Caki-1 and 786-O cells transfected with circPDHK1 siRNA and/or PPP1CA siRNA. Bars = 200 μm. E–G Transwell migration and invasion assays to detect cell migration ability of Caki-1 and 786-O cells transfected with the control siRNA, a circPDHK1 siRNA, a PPP1CA knockdown siRNA and cotransfected with the circPDHK1 siRNA and the PPP1CA knockdown siRNA. Bars = 100 μm. H Western blot detection of the phosphorylation levels of the AKT-mTOR signaling pathway in Caki-1 and 786 cells. I Analysis of subcutaneous tumors in mice injected with cholesterol-modified the control siRNA, a circPDHK1 siRNA and cotransfected with the circPDHK1 siRNA and the PPP1CA knockdown siRNA. Tumor volumes calculated every 2 d. J At Day 37 after treatment, all mice were sacrificed, and subcutaneous tumors were dissected and recorded. Tumor weights in each group as indicated. K Images of subcutaneous tumors. (n = 6, each group). L, M Representative photographs and quantification of Ki-67 IHC staining in subcutaneous tumors. *P < 0.05; **P < 0.01; ***P < 0.001; ns, no significance