a,b, Bulk RNA-seq analysis of HUVEC (EC), HASMC (SMC), CD4+ effector memory T cell (TEM) and macrophage (M) gene expression before and after stimulation with TGF-β1 (a) and inflammatory gene expression (b). Each lane is an average of two biologically independent samples. c, ChIP-seq analysis of canonical TGF-β signalling in ECs and SMCs. Note increased SMAD2/3 binding to regulatory elements of CCL2, CLDN5 and SERPINE1 in ECs but not SMCs. Pol II S2 and Pol II S5 ChIP in combination with bulk RNA-seq indicate increased expression of CCL2 and SERPINE1 and decreased expression of CLDN5 after TGF-β1 stimulation; n = 1 per experimental condition. d, Relative CLDN5 expression in primary ECs isolated from control (Cdh5-CreERT2;Tgfbr1fl/fl;Tgfbr2fl/fl;mT/mG mice treated with corn oil) and TGF-βRiEC (Cdh5-CreERT2;Tgfbr1fl/fl;Tgfbr2fl/fl;mT/mG mice treated with tamoxifen) mice. Bars throughout indicate the mean ± s.e.m. 0 h, P = 0.004; 6 h, P = 0.001. Two-tailed Student’s t-tests (n = 3 animals per group for 0 h, n = 6 animals per group for 6 h). e, Intravenous Evans blue dye administration following a subcutaneous TNF-α injection into mouse ear. Scale bar, 2 mm; 0 min, P = 0.56; 15 min, P = 0.000066, two-tailed Student’s t-tests (n = 6 animals per group for 0 min, n = 7 animals per group for 15 min). f,g, Mouse ear sections were stained with anti-ICAM, VCAM-1, Ly6G and CD45 antibodies 6 h and 3 days following a subcutaneous TNF-α injection. Note a strong reduction in expression of both adhesion molecules (f) and reduced presence of inflammatory cells (g) in TGF-βRiEC mice. Scale bar, 16 μm; n = 7 animals per group. ICAM1+ area 6 h, P = 0.001; ICAM1+ area 3 d, P = 0.0002; VCAM-1+ area 6 h, P = 0.0001; VCAM-1+ area 3 d, P = 0.00001; Ly6G+ cells 6 h, P = 0.0499; Ly6G+ cells 3 d, P = 0.0004; CD45+ cells 6 h, P = 0.051; CD45+ cells 3 d, P = 0.00004. Two-tailed Student’s t-tests.