Figure 3.

Vasculostatin-120 (Vstat120) is secreted by OvCa cells infected with 34.5ENVE, and conditioned media from 34.5ENVE-infected OvCa reduces endothelial cell migration in vitro. (A) Immunoblot of the indicated OvCa cell lysates harvested 24 hours after infection with 34.5ENVE (“ENVE;” at MOI=0.1), revertant ENVE (“Rev;” at MOI=0.1), or PBS. Blots were probed for expression of VStat120 (indicated with arrow), viral infected cell protein 4 (ICP4), and cellular GAPDH. Ladder indicates position. (B) Immunoblot of concentrated conditioned media from SKOV3. ip1 or patient ascites-derived tumor cells (number 140), harvested and concentrated 12 hours after infection with 34.5ENVE (“ENVE;” MOI= 2), revertant ENVE (“Rev;” MOI= 2), or PBS. Blot was probed for Vstat120 (indicated with arrow). Ladder indicates position of molecular weight markers.(C) Migration of human dermal microvascular endothelial cells (HDMEC) treated with concentrated conditioned media (CM) collected from OvCa cells infected with 34.5ENVE, revertant ENVE or PBS for 12 hours. Media from infected SKOV3 or patient ascites-derived tumor cells (number 140) was collected and concentrated, and added to bottom chamber of a transwell assay; HDMEC were seeded on top of the membrane in the upper chamber. HDMEC migration was assessed by quantifying number of cells that had migrated through the membrane pores 12 hours after seeding. Data shown are mean number of cells migrated though membrane relative to control; error bars indicate standard deviation. ANOVA models used to compare the mean of multiple groups; p value adjusted by Holm’s procedure.