Fig. 6

CAF-derived IL-1β upregulates expression of EC adhesion molecules and invasiveness markers. a, b C166 endothelial cells were incubated for 24 h in control medium, medium containing recombinant IL-1β (5 ng/mL), conditioned medium of MSU-treated WT mammary fibroblasts (Balb/C), or conditioned medium of MSU-treated Il1b^−/− fibroblasts. The expression of genes involved in tumour-cell intravasation/extravasation was analysed using qRT-PCR. Representative of three independent experiments. c, d Trans-endothelial migration of mCherry-labelled Met-1 cells supplemented with recombinant IL-1β (5 ng/mL), conditioned medium of WT NMFs, conditioned medium of Il1b^−/− NMFs, or control medium. c Representative images of trans-migrated Met1-mcherry cells. Scale bar, 100 μm. d Quantification of migrating cells per field of view. Representative of two independent experiments. e, f qRT-PCR analysis of the expression of adhesion and migration-related genes in endothelial cells (CD31⁺CD45⁻EpCAM⁻) isolated by FACS from e primary 4T1 tumours or f lungs of tumour-bearing mice. n = pool of six mice per group. Representative of two independent biological experiments. g qRT-PCR analysis of the expression of invasion-related genes in tumour cells (EpCAM⁺CD45⁻) isolated by FACS from 4T1 mammary tumours. n = pool of six tumours per group. Results are representative of two independent experiments. In a, b, d data are presented as mean ± s.d. of biological repeats; One-way analysis of variance test followed by Tukey’s multiple comparisons test. In e–g data are presented as mean ± s.d. of technical repeats; Welch’s t-test. Source data are provided as a Source Data file