Fig. 3

RDM1 loss suppresses cell cycle, promotes apoptosis, and suppresses tumor development in vivo. (A–D) After shNC or shRDM1 treatment, the cell cycle progression of 786-O and Caki-1 cells could be observed through assays. Student´s t‑test. Error bars, mean ± SD. ***p < 0.001. (E) Western blotting results showing Cyclin D1, CDK4, and CDK6 protein levels before and after shRNA transfection. (F and G) PI and Annexin V staining of shNC- or shRDM1-treated human ccRCC cells, and results analyzed using flow cytometry. Student´s t‑test. Error bars, mean ± SD. ***p < 0.001. (H) Western blotting results showing Bax, Bcl-2, and Caspase 3 protein levels before and after shRNA transfection. (I) Excision and imaging of the mice xenotransplanted with human 786-O cells transfected with shNC or shRDM1. (J) Tumor volume and weight (K) at the end points of subcutaneous xenograft tumors (n = 10 for every group). Student´s t-test. Error bars, mean ± SD. **p < 0.01. (L-M) Representative images of the TUNEL assay. Scale bars, 20 μm. Student´s t‑test. Error bars, mean ± SD. ***p < 0.001. (N) Representative images of IHC. Scale bars, 100 μm.