Supplementary Figure 4.

Effect of long-term combinatorial silencing of early and middle Ad5 genes on Ad5 replication. A549 cells were transfected with siRNAs directed against the E1A, DNA polymerase (Pol), pTP, and IVa2 genes, or a non-targeting control siRNA (neg. ctrl.), either alone (at a concentration of 10 nM) or in combination (at a concentration of 5 nM each). At 24 h after transfection, cells were infected with Ad5 at an MOI of 0.01 TCID₅₀/cell. Cells and supernatants from triplicate infection experiments were harvested at 6 days post-infection, and virus genome copy numbers (mean ± SD; n = 3) were determined by qPCR, using E1A-specific primers. For each smple, real-time qPCR quantification was performed in duplicate. ^**p < 0.01.