Fig. 1

The characterization of human colorectal cancer-derived primary CAFs and NFs. a Immunofluorescence of FSP and α-SMA of CAFs and NFs (magnification, ×200). CC-CAFs were positive for the expression of FSP, α-SMA (b) the protein expression of CAFs and NFs was determined by western-blot. c The mRNA expression of FAP, α-SMA, vimentin was analyzed by Q-PCR, GAPDH serves as internal control. Error bars represent mean ± s.d; *P < 0.05, by unpaired two-sided Student’s t-test. d mRNA expression of hTERT was analyzed by Q-PCR, GAPDH serves as internal control (n = 3). e Expression of CD44 in liver metastasis site and primary tumor site (magnification, ×200). f The protein expression of liver metastasis site and primary tumor site was evaluated by western-blot (LM liver metastasis site, PT primary tumor site). g, i Liver metastasis site-derived CRC cells in orthotopic liver metastatic model was isolated, and expression of CD44 was determined by western-blot, and CD44 expression in GFP(+) subpopulation was analyzed by flow cytometry. h CD44 expression of primary tumor and CT26-GFP was evaluated by western blot and representative results from one of the three independent experiments are presented. Error bars represent mean ± s.d; *P < 0.05, **P < 0.01; ***P < 0.001; ****P < 0.0001, n.s not significant; by one-way analysis of variance (ANOVA)