Fig. 6

In vivo allograft tumor growth of RASV12-transformed MEF expressing (Wt RAS MEF) or not (Vdac1 ^−/− RAS MEF) VDAC1. a Wt RAS (+) and Vdac1 ^−/− RAS (−) MEF were incubated in Nx for 24 h and cell lysates were analyzed by immunoblotting for RAS and VDAC. β-tubulin was used as a control. b Allograft tumor growth of Wt RAS MEF and Vdac1 ^−/− RAS MEF injected into one of the flanks of nod-scid mice. Five mice were studied per condition. A p < 0.0000001 shows significant differences. c Wt RAS (tumors 1, 2, and 3) and Vdac1 ^−/− RAS MEF-derived tumors (tumors 4, 5, and 6) cell lysates were analyzed by immunoblotting for HIF-1α, HIF-2α, and VDAC. β-tubulin was used as a loading control. d Immunofluorescence of CD31 to detect blood vessels in Wt RAS and Vdac1 ^−/− RAS MEF-derived tumors. e Immunofluorescence of α-smooth muscle actin to visualize the structure of blood vessels in Wt RAS and Vdac1 ^−/− RAS MEF-derived tumors. f Representative images of Masson’s trichrome stained Wt RAS and Vdac1 ^−/− RAS MEF-derived tumors. Red staining indicates red blood cells. g Expression of the mRNA of Mmp3, Col18A1 (Endostatin) and Thbd in Wt RAS (Wt RAS MEF) and Vdac1 ^−/− RAS MEF-derived tumors (Vdac1 ^−/− RAS MEF). h Expression of the pro-inflammatory cytokine mRNA of Il8 and Cxcl5 in Wt RAS (Wt RAS MEF) and Vdac1 ^−/− RAS MEF-derived tumors (Vdac1 ^−/− RAS MEF). i Expression of the mRNA of Cdkn2a in Wt RAS (Wt RAS MEF) and Vdac1 ^−/− RAS MEF-derived tumors (Vdac1 ^−/− RAS MEF)