Selective replication and oncolysis of Ad.hTERT-E1A-TK. A. Comparison of cytopathic effects between NSCLC NCIH460 and primary fibroblasts. NSCLC NCIH460 and primary fibroblasts were plated into 6-well plates and infected with 10 MOI of Ad.hTERT-E1A-TK, dl309 or Ad.GFP. 5 days later cytopathic effects were observed and photographed by light microscopy. B. The virus progeny production in NCIH460 cells and primary fibroblasts. NCIH460 and primary fibroblasts were infected with 10 MOI of Ad.hTERT-E1A-TK for 4 h then washed once with PBS and then cultured with fresh medium. On 24 h and day 5 post infection, the cells were harvested for plaque assay. The plaques on HEK293 cells were counted and plotted. C. Western blotting analysis of E1A gene expression. NCIH460 and SW1990 Cells were infected with Ad-hTERT-E1A-TK at a MOI of 10. Cell lysates were harvested 48 h later, and immunobloted by anti E1A antibody. HEK293 cells which had been transfected with E1A were used as positive control, and uninfected NCIH460 cells were used as negative control.