Figure 3
From: DNA intercalator stimulates influenza transcription and virus replication
Effects of ActD and DRB on RNAPII phosphorylation and viral transcription. Prior to infection, MDCK cells were treated with ActD or DRB in MEM supplemented with 10% foetal calf serum and 1% penicillin/streptomycin for three hours, followed by infection with A/WSN/33 (MOI = 2) in the presence of the corresponding drugs. The infected cells were cultured in a medium containing ActD or DRB, and culture supernatants were harvested at six hours post-infection. (A) Effects of ActD and DRB on RNAPII phosphorylation in infected cells. Total cell lysates were harvested and studied by western blot analysis using anti-RNAPII (N20) and anti-β actin (C4) antibodies (Santa Cruz Biotechnology, Santa Cruz, USA). The signals for RNAPIIo and RNAPIIa are indicated. Beta-actin was used as the loading control. (B) Effects of ActD and DRB on viral RNA synthesis. Total RNA was analyzed with primer extension assays, as described previously (10). The bands representing the mRNA, cRNA and vRNA of segment 5 are marked by arrows.