Figure 5.

GPS491 alters adenovirus RNA expression/processing and inhibits adenovirus DNA amplification. A549 cells were infected with HAdV-C5 at an input MOI of 100 IU/cell for 1 h after which virus inoculum was removed and replaced with media containing DMSO or GPS491 (2.5 or 5 µM). (a) Total RNA was extracted at 8 h, 16 h, or 24 h after virus infection. After cDNA generation, RT-qPCR was performed using primers for E1A, E1B, E2A, E2B, and E4 RNAs (Supplementary Table S1). Values are expressed relative to RNA abundance observed 24 h p.i. in the presence of 1% DMSO. Shown are data from samples treated with 2.5 µM GPS491. (b) The effect of GPS491 on E1A RNA processing. Shown on the top is the schematic diagram of E1A RNA processing indicating the major E1A mRNA isoforms generated by alternative splicing. In the middle is a representative gel of the E1A RNA amplicons generated from cDNA. The graph on the bottom represents quantitation of amplicons generated across n > 3 independent assays. (c) At 16 h, 20 h, and 24 h p.i., total DNA was isolated from cells treated with DMSO or 2.5 µM GPS491 and levels of adenoviral DNA were determined by qPCR. Data are indicated as mean ± SD, * p ≤ 0.05, ** p ≤ 0.01, and *** p ≤ 0.001.