Extended Data Figure 2: TCs induce necroptosis in ECs in vivo.
From: Tumour-cell-induced endothelial cell necroptosis via death receptor 6 promotes metastasis
a, Quantitative evaluation of cleaved caspase-3- and EthD-III-positive cells in lungs of C57BL/6 WT animals at the indicated time points after i.v. injection of B16 TCs on the basis of the analysis of confocal microscopy images as shown in main Fig. 2a, b. TCs were injected at the same time and lung isolation occurred at the indicated time points. Injection of PBS served as control. b, Quantification of EthD-III-positive ECs in lungs of WT animals 6 h after i.v. injection of B16 or LLC1 TCs. c–e, Representative confocal images of lung sections taken 6 h after i.v. injection of B16 or LLC1 TCs (c, d) or injection of equal amounts of fluorescently labelled 15 μm microspheres (e) into WT animals and stained for the indicated markers. Arrowheads in c indicate EthD-III-positive cells. Isolated lungs from animals cultured ex vivo in the presence of staurosporine served as positive controls in d. Scale bar, 20 μm. Shown are representative data of two (a) or three (b) independent experiments with mean values ± s.e.m. from n = 3 animals per time point (a) or n = 6 animals per group (b). **P < 0.01; ***P < 0.001. One-way ANOVA and Bonferroni’s post hoc test.
