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. 2014 Apr 19;105(5):560–567. doi: 10.1111/cas.12394

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© 2014 The Authors. Cancer Science published by Wiley Publishing Asia Pty Ltd on behalf of Japanese Cancer Association.

This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

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Functional analysis of tumor endothelial cell (TEC) markers using siRNA-mediated gene silencing. Melanoma-EC were transfected using three different siRNA for each gene. Silencing of each TEC marker 48 h after transfection was evaluated using qRT-PCR (upper panel). Cell proliferation was measured after 72 h using Alamar Blue (lower panel). Cell migration towards the chemoattractant EGM-2MV for 20 h was evaluated using the BD BioCoat Angiogenesis System: Endothelial Cell Migration (BD Biosciences) (lower panel). siRNA-mediated effects of each TEC marker are shown in (a–e) (**P < 0.01; *P < 0.05). (a) Tmem176b; (b) Pcdhb22; (c) Nsg1; (d) Enah; and (e) Def6. NC, Negative control siRNA.