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. 2019 Jul 24;19:192. doi: 10.1186/s12935-019-0914-y

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© The Author(s) 2019

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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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)