Extended Data Fig. 6: CAFs promote metalloproteinase activity in CRC cells.
From: Patient-derived mini-colons enable long-term modeling of tumor–microenvironment complexity
a, Western blot showing the expression of the indicated proteins in CRC organoids in control and CAF-conditioned medium conditions. b, Western blot-based quantitation of MAPK1/3 (ERK1/2) phosphorylation in CRC organoids in control and CAF-conditioned medium conditions. ***P < 0.0001 (two-tailed unpaired t test; n = 6 for each group). c,d, Quantitation of the invasive front area (c) and epithelium thickness (d) in CRC mini-colons co-cultured with autologous CAFs and treated with the indicated compounds. *P = 0.0357; **P = 0.0015 (c), 0.0096 (d); ***P < 0.0001 (two-way ANOVA and Sidak’s multiple comparisons test; n = 3 mini-colons (c) and 18 inter-crypt epithelium sections (d) for each group). e, Brightfield images of CAF invasion of the stromal hydrogel when CRC mini-colons are treated with the indicated compounds. The maximum reach is indicated with an arrow. Scale bar, 150 μm. f, Quantitation of CAF invasion of the stromal hydrogel 7 days after CRC mini-colons are treated with the indicated compounds. ***P = 0.0002 (two-tailed unpaired t test; n = 6 for each group). g, qRT-PCR based quantitation of MMP7 mRNA in the indicated knockdown cell lines. ***P < 0.0001 (one-way ANOVA and Tukey’s multiple comparisons test; n = 3 for each group). h, Quantitation of the invasive front reach in the indicated CRC mini-colon knockdowns co-cultured with autologous CAFs. **P = 0.0012 (day 7), 0.0093 (day 15); ***P < 0.0001 (two-way ANOVA and Sidak’s multiple comparisons test; n = 24 invasive fronts for each group). In b-d and f-h, data represent mean ± SEM. Ctrl, control; CM, conditioned medium; KDa, kilodalton. In all panels, data refer to patient #MS.
