Fig. 5: Endothelial structures formed within the cancerous CAF stroma.

Number of endothelial structures formed within a HT29 and b HCT116 tumouroid stromal compartments at day 21 (mean ± SEM) containing either HDF or one of six patient-specific CAF tissue samples. All n = 3 with four technical repeats and showing Kruskal–Wallis with Dunn’s multiple-comparison test P values, with values 0.05 = *, 0.005 = **, 0.0005 = *** and 0.00005 = ****. c Example images of normal endothelial structure formation within a HDF-containing stroma HCT116 tumouroid at day 21 at the cancer-stromal edge and with a d budded invasive body within a HT29 tumouroid. Scale bar = 100 µm for the left and 50 µm for the right image, and red = CK20, green = CD31 and blue = DAPI. e Images showing the decreased formation of complex endothelial structures within a CAF-containing stroma at day 21 near the cancer-stroma edge, f as well as around a budding invasive body, the only occurrence of endothelial structures within these conditions. Scale bar = 50 µm and red = CK20, green = CD31 and blue = DAPI. Active VE-Cadherin protein released into the media of one of six CAF-containing g HT29 or h HCT116 tumouroids over 21 days (mean ± SEM). Paired t-test comparison test between day 2 and day 21 with P values 0.05 = *, 0.005 = **, 0.0005 = *** and 0.00005 = **** with DOF = 2 for both and t value for HT29 = 8.115 and HCT116 = 4.766. i FBLN5 (fibulin-5) expression at day 21 within the HDF or one of six CAF-containing HT29 or HCT116 tumouroids (mean ± SEM). The value shown is normalised to HPRT1 mRNA levels (mean ± SEM) with n = 3 and 3 technical repeats. Ordinary one-way ANOVA Dunnett’s multiple-comparison test with P values 0.05 = *, 0.005 = **, 0.0005 = *** and 0.00005 = ****.