Fig. 8

Analysis of cytotoxic CRAd effects combined with Avastin. a) The monolayers SKOV3ip.1, OV-4, and OVCAR3 OvCa cells were infected with each CRAd alone or two CRAd vectors together at MOI of 10 vp/cell. Avastin was used to supplement infection and mock infection medium at the indicated concentrations (μg/ml). The infected and uninfected cells were subjected to CellTox™ assay (Promega) by adding DNA-binding cyanine dye on day 3 and monitoring the increase in fluorescent signal intensity, which is proportional to cytotoxicity till day 5 post-infection to detect any cell killing effects. The cells in 96-well plates were red using the Synergy-HT plate reader with 485 nm excitation and 520 nm emission filters. The data represent the mean values of relative fluorescent units (RFU) detected for each Avastin concentration after subtracting background signal detected with cyanine dye added but no virus or Avastin. b) The Cell Proliferation Assay (Promega) was carried out 6 days post-infection to detect cells that stayed alive after exposure to CRAd and/or Avastin. The plates were red using the Synergy-HT plate reader set at 490 nm. The mean values of optical density (OD) detected for each Avastin concentration are presented after subtracting background signal detected in monolayers that were not treated with virus or Avastin. Each data point represents the cumulative mean ± SD (error bars are smaller than the symbols, p ≤ 0.05)