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. 2013 Dec 23;8(12):e83933. doi: 10.1371/journal.pone.0083933

Figure 1. Endogenous ROBO4 upregulation despite lower vascular density in orthotopic and xenograft tumors.

Figure 1

A. Immunofluorescence of the vascular endothelium in liver (upper panel) and 786-O human renal cell carcinoma (RCC) subcutaneous xenograft tumor (lower panel). B. Vascular area analysis of liver (Li), kidney orthotopic (KO) tumors, and subcutaneous (SC) xenograft tumors (n = 6 mice analyzed). C. Immunoblot of endogenous ROBO4 and the endothelial cell specific VE-cadherin from liver, kidney orthotopic and subcutaneous xenograft 786-O RCC tumors, and from the derivative 786-O cells grown in culture. D. Densitometry analysis of VE-cadherin/tubulin ratio from C mirrors the vascular area determination in B. E. Densitometry analysis of endogenous ROBO4 normalized to VE-cadherin expression reveals a 1.4- to 2-fold increase in SC and KO tumors compared to liver. C–E: Immunoblot and densitometry was repeated twice with two independent sets of protein extracts from two different tumor-bearing mice with essentially the same results. A. Magnification: 100X, Red: endomucin/CD31 antibody cocktail, Blue: DAPI. B. *p<0.05, one way ANOVA with Tukey's correction, mean ± SD.