Figure 4

Analysis of CRAd-hsp70E1Δ55k replication using real-time PCR. a, to confirm E1B55k deletion from CRAd-hsp70E1Δ55k and absence of wild-type Ad, 150,000 A549 cells were plated in 12-well plates and infected after 24-hrs in triplicates with 10 MOI (iu) of CRAd hsp70E1Δ55k, AdCMVE1 or Ad5luc3. Thirty-six hours later cellular DNA was extracted and real-time PCR was performed to measure the E1B gene copy number using primers encompassing the E1B19k (forward primer) and E1B55k (reverse primer) sequences. b, to evaluate CRAd-hsp70E1Δ55k replication relative to non-attenuated Ad, real-time PCR was performed on DNA extracted from cells infected above in (a), and the E4 gene copy number was measured. *, p < 0.05 for attenuated CRAd-hsp70E1Δ55k replication vs. Ad5luc3 (t test).