Figure 1.

Induction of apoptosis by oncolytic NDV/FMW in lung cancer cells. A. H460 cells were infected with or without (mock-infected) NDV/FMW (MOI = 1) for the indicated time-points. Cells at 24 and 48 h post-infection (hpi) were double-stained with Annexin V and propidium iodide (PI) and analyzed by flow cytometry. The cell population in the right lower quadrant (PI-negative, Annexin V-positive) and the right upper quadrant (Annexin V/PI positive) are represented. Data shown are representative of three independent experiments (***P<0.001). B. Cells at 12, 24 and 48 hpi were lysed and activation of caspase-3 and cleaved poly (ADP-ribose) polymerase (PARP) was examined by immunoblot analysis (n = 2). Replication of NDV/FMW was detected by examination of the expression of hemagglutinin-neuraminidase protein (HN) protein. To control for loading, a-tubulin was also used. Immunoblots shown are representative of two independent experiments. C. Cells were pre-treated with either Z-VAD-FMK (Z-VAD, 100 μM) or Necrostain-1 (Nec-1, 20 μM) or mock-treated, following infection of NDV/FMW for 48 h. Apoptosis was analyzed by flow cytometry. Data are representative of three independent experiments (**P<0.01, n.s = not significant).