Fig. 3. CRC patient serum-derived TIMP1^EV promotes ECM remodelling.

A Representative bright field images of collagen matrigel lattices with embedded pFs treated with CRC EVs (HCT116, HT29, SW620) after 48 h (n = 3), white dashed line highlighting the matrix margins. B Percentage of contraction shown in (A) exerted by SFM, CM and EV treated pFs as calculated using imageJ. C Representative bright field images of collagen matrigel lattices with embedded pFs treated with EVs from TIMP1^OE, TIMP1^WT, and TIMP1^KO cell lines after 48 h, white dashed line highlighting the matrix margins. SFM treated pFs were used as control (n = 5). D Percentage of matrix contraction shown in (C) calculated using imageJ (n = 5). E TIMP-1 mRNA expression in pFs treated with serum EVs from HD (n = 20), CRC liver MET (n = 46) and CRC (n = 18) using qPCR. Values were normalised to mean CM. F Representative Immunoblot analysis of TIMP1 expression in pFs treated with serum EVs from HD, CRC liver MET and CRC. (Shown: n = 3, Total: n = 27 per group). B-actin (ACTB) was used as a loading control. G Densitometry quantification of TIMP1 expression normalised to ACTB of all blots shown in (F) using imageJ (n = 27). H Representative bright field images of matrix contraction of pFs treated with serum EVs from HD, CRC liver MET, and CRC. (Shown: n = 3, Total: n = 12 per group). I Percentage of contraction shown in (H) of all samples calculated using imageJ (n = 12). Error bars depict mean ± SEM. P values were calculated by unpaired t-test except for (B) one-way ANOVA test was used. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns not significant.