Fig. 4

KPC1 p.M8V editing confers loss-of-function effects on the tumor suppressive roles of wide-type KPC1 in iCCAs. a-d The effects of overexpression of wide-type KPC1 (KPC1-WT) or edited KPC1 at p.M8V (A22G) (KPC1-EDT) on cells growth (A), plate colony formation (B), migration (C) and invasion (D) in QBC939 or RBE cells. e The effect of overexpression of KPC1-WT or KPC1-EDT on subcutaneous tumor growth (assessed by tumor volume) in nude mice. RBE cells with empty vector, overexpression of KPC1-WT or KPC1-EDT were implanted subcutaneously at the back of the mice (n = 9 mice/group). f Weights of the xenografted tumors at the 28th day after subcutaneous implantation at the back of the mice from the indicated groups. g The effect of overexpression of KPC1-WT or KPC1-EDT on tumor cells proliferation activities in nude mice, which was indicated by immunohistochemistry (IHC) staining for Ki-67. Left panel, the representative hematoxylin–eosin (H&E) and Ki-67 IHC staining images for the xenografted tumors. h–k The effect of overexpression of KPC1-WT or KPC1-EDT on tumor metastasis in nude mice. The empty vector-, KPC1-WT- or KPC1-EDT-overexpressed RBE cells (each also expressing luciferase) were injected via the tail vein into the nude mice (n = 6 mice/group). (H) Representative bioluminescence (BLI) images of the mice injected with RBE cells via the tail veins. (I) Kaplan–Meier plot of mice. The P value was assessed by log-rank test. (J) Representative ex vivo BLI images of metastases in lungs. (K) H&E staining of metastases in lungs. Scale bar, 100 µm. Data are presented as the mean ± standard deviation (s.d.). ^*P < 0.05, ^**P < 0.01 and ^***P < 0.001; assessed by Student’s t test