Fig 5.
Influence of 55R E1A on viral gene expression. (A to D) Contact-inhibited primary IMR-90 fibroblasts were infected with dl309, JM17-55R, dl521, or dl312 at an MOI of 5. Cells were collected at 24, 48, 72, 96, or 120 hpi. Samples were subjected to qRT-PCR analysis using primers specific to transcripts controlled by various HAdV promoters, including E1B (A), E3A (B), E4orf6/7 (C), and hexon (D). (E) Relative expression levels at 144 hpi of viral transcripts from cells infected with dl309, JM17-55R, dl521, or dl312. Samples were treated with DNase I to remove contaminating viral DNA, and no-RT controls from 96 hpi were run with each sample to ensure the efficiency of this process. All samples were normalized internally to GAPDH (A to E) and then to the levels of gene expression observed in dl312-infected cells (A to D), using the ΔΔCT method.