Figure 5
From: Vasohibin inhibits angiogenic sprouting in vitro and supports vascular maturation processes in vivo
Effects of stable overexpression of mVASH1 on tumor growth and large vessel growth. B16F10 cell clones with mVASH1 overexpression (VH#1, 2, 4) or control clones (co#10, 11, 12) were subcutaneously transplanted in C57BL/6 mice and melanomas removed after 14 days. (A) VASH1 gene expression was analyzed in all tumor samples by RT-PCR. Actin beta (ACTB) served as housekeeping gene. (B) Statistical analysis of large vessels (diameter>50 μm) in controls and mVASH1-overexpressing tumors. (C) Analysis of vasular area in controls and mVASH1-overexpressing tumors (D) Western blot analysis of tissue protein extracts of a VASH1 overexpressing and control melanoma. Immunoreactive VASH1 protein was present in high amounts and also carbonic anhydrase 9 (CA9) was higher expressed in VASH1 overexpressing melanomas. VEGF protein was not altered. Tubulin alpha served as internal loading control. (E) Proangiogenic gene expression was analyzed by qPCR in controls and mVASH1-overexpressing tumors. Proangiogenic gene expression of VASH1 overexpressing tumors (n = 9) was quantified relative to the mean of all control tumors (n = 9). Stars indicate p values < 0.05 and bars mean ± SEM.