A3B knockdown promotes replication of strain A12.

A549 lung cancer cells were transduced to stably express APOBEC3B-targeting shRNAs (shA3B) or a nontargeting control shRNA (scramble). (A) A3B mRNA levels were quantified by RTqPCR and expressed relative to GAPDH mRNA. (B) A3B protein levels were assessed by western blot, quantified by densitometry and normalized with HSP90. (C) Deamination activities of the cell lysates were expressed as the percentage of substrate converted into product and compared to the deamination activity of the WT cells. P-values were calculated by t-tests. (D-G) A549-WT, -scramble and -shA3B were infected with HAdV-A12, -B3 or -C2 at a MOI = 0.03 and analyzed at different time points post infection. (D-E) The percentage of infected cells were quantified by flow cytometry. The results of the first replicate are shown in panel D and the data from the 3 replicates are shown in panel E. Intracellular viral DNA levels (F) and extracellular viral DNA levels (G) were quantified by qPCR. P-values were calculated by ratio t-tests. Only significant differences are shown.