Figure 7: Functional significance of VE-cadherin expression for SCLC VM in vitro and in vivo.

(a) Western blot analysis of VE-cadherin expression levels in VM proficient C8161 melanoma cells and H446, H446 non-silencing empty vector controls, H446 VE-cadherin shRNA knockdown and H1048 SCLC cells. (b) VM-like network formation on matrigel for VE-cadherin^+VE cells (C8161, H446 and H446 E.V) and VE-cadherin^−VE cells (H1048 and H446 VE-cadherin KD) lacking network formation. Representative images are shown for n>3 experiments. Scale bar, 200 μm. (c) Left panel, representative images of anti-human anti-VE-cadherin staining in H446 parental and H446 VE-cadherin KD xenografts. Scale bar, 50 μm. (c) Right panel percentage of cells positive for VE-cadherin in H446 and H446 VE-cadherin KD xenografts (***P=0.0001, two-tailed t-test), n=10 animals per group. (d) Western blot analysis VE-cadherin expression in H446 and H446 VE-cadherin KD xenografts (n=6 tumours for each group). (e) Left panel, representative images of anti-mouse anti-CD31/PAS staining in H446 Parental and H446 VE-cadherin KD tumours. Scale bars, 50 μm. (e) Right panel VM ratio in H446 parental (n=10) and H446 VE-cadherin KD xenografts (n=8; ***P=0.0005, two-tailed Mann–Whitney’s test). (f) Tumour growth rates in H446 (black) and H446 VE-cadherin KD (grey) tumours (n=10 animals per group). (g) Days to 200 mm³ volume tumours in H446 and H446 VE-cadherin xenografts KD (n=20 animals per group, (***P<0.0001, two-tailed t-test). Error bars show s.e.m.