Establishment of CAFs. (A) Rosa26-mT mouse treated with DEN for 17 weeks, and waiting 30 weeks for tumor formation. Then mouse CAFs were cultured according to our protocol. (B) Representative immunohistochemistry staining of α-SMA in mouse and human primary tissue (magnification, 400×). (C) Representative image of established human and mouse CAFs (magnification, 100×). (D) Representative immunofluorescence staining of α-SMA, FAP, EpCAM, AFP, CD45, and CD31 in mouse and human CAFs (magnification, 400×). DAPI, 4′,6-diamidino-2-phenylindole; hCAF, human cancer associated fibroblast; mCAF, mouse cancer associated fibroblast; mT, membrane tomato; RFP, red fluorescent protein.