Fig. 4: Pericyte FAK deficiency stimulates increased levels of Tissue Factor expression in B16F0 cells.

a Cytokine array. Cyr61 quantitation in FAK^KO pericytes. Chart represents mean ± s.e.m. Representative images of Cyr61dots on array. n = 3 biological repeats in duplicate. ***p = 0.0005. This result was confirmed by western blotting of Cyr61 with the same lysates. Chart represents mean Cyr61 densitometry, n = 2 experimental repeats. b Western blot. Cyr61 expression is increased in FAK^KO pericytes. Depletion of Axl (FAK-null;Axl^KO) abrogated Cyr61 expression. Chart represents mean ± s.e.m., n = 3 biological repeats. *p = 0.0213, **p = 0.009. c Recombinant Cyr61 stimulated endothelial cell spheroid sprouting. Chart represents mean ± s.e.m., ****p < 0.0001. Scale bar, 100 μm. n = 37 PBS treated spheroids and 30 Cyr61 treated spheroids. d Western blot. Cyr61-depletion in FAK-null;Cyr61^KO pericytes. Chart represents mean Cyr61 levels ± s.e.m., *p = 0.0133; n = 3 experimental repeats. e Tumour growth. Line graph represents mean ± s.e.m., **p = 0.0046, ****p < 0.0001. FAK-null;Cyr61^KO: n = 7 mice, FAK-null;Cas9: n =  5 mice, WT;Cas9: n = 10 mice. Tumour blood vessel density. Chart represents mean blood vessel density/mm² ± s.e.m., ***p = 0.0006. n = 28, 30 and 14 fields of view from 4 FAK-null;Cas9 tumours; n = 4 FAK-null;Cyr61^KO tumours, and n = 3 WTCas9 tumour sections respectively. Scale bar, 50 μm. Immunostaining for endomucin. f Western blot. p-AKT expression in WT and FAK^KO pericytes. Line graph shows mean ± s.e.m., **p = 0.0043; n = 3 experimental repeats. g Western blot. PI3-kinase inhibitor (GDC-0941) reduced expression of Cyr61 after stimulation with Gas6 in FAK^KO pericytes (10 and 20 min). Chart represent mean ± s.e.m Cyr61 levels. **p = 0.0047; n = 3 biological repeats. h Cytokine array. Tissue factor (TF) quantification. Representative images of dot blots. Chart represents mean ± s.e.m.TF levels, n = 4 biological repeats. *p = 0.0295. Western blot analysis confirmed increased TF production in B16F0 melanoma cells co-cultured with FAK^KO pericytes. i Western blotting. B16F0 cells had increased expression of TF in response to exogenous Cyr61 (10 μg/ml). Chart represents mean ± s.e.m. n = 3 experimental repeats. **p = 0.0048. j Western blot. TF expression is reduced after TF depletion in B16F0 cells. Chart represents mean ± s.e.m., n = 3 lysates. **p = 0.0016. Tumour volume measurements. Chart represents mean ± s.e.m., *p = 0.0453; n = 10 pdgfrβcre+;fakfl/fl and 16 pdgfrβcre-;fakfl/fl mice. a–d, f, g–j two-sided Students t test. e, j one-way ANOVA; ns not significant. Source data are provided as a Source Data file.