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. 2015 May 20;2:15008. doi: 10.1038/mto.2015.8

Figure 1.

Figure 1

RAMBO is antiangiogenic and cytotoxic toward human squamous cell carcinoma (SCC) cells. (a) Western blot analysis of CAL27 and UM-SCC-74A cells treated with PBS, rHSVQ1, or RAMBO. The cells were harvested at 48 hours postinfection and analyzed for expression of Vstat120. GAPDH was used as a loading control. Note the presence of Vstat120 in cells infected with RAMBO. Data are representative results from a total of three separate experiments. (b,c) Antiangiogenic effect of RAMBO in CAL27 xenografts tumors. (b) Representative images of tumor sections illustrating microvessel density assessment stained with CD31 antibody. Arrows are representative vessels from each group (n = 3/group). (c) Quantification of CD31 immunostained vessels performed. Data shown are average vessel density ± SD (P < 0.05). (d,e) SCC cells are sensitive to RAMBO in vitro. (d) Quantification of cell viability of six different SCC cell lines infected with RAMBO relative to uninfected cells at the indicated multiplicity of infection (MOI) 48 hours postinfection. Data shown are % cell viability relative to uninfected cells and representative results from a total of three separate experiments. Data were analyzed by the paired student’s t-test and error bars indicate ± SD for each group. (e) Representative images of UM-SCC-74A cells 48 hours after RAMBO infection at the indicated MOI. Note the rounded morphology and reduced attachment of infected cells 48 hours postincubation.