Fig 9.

Expression of PARP12L strongly affects expression of genes carried by the recombinant VEEV genome. (A) Schematic representation of VEEV replicons encoding PARP12L in either the direct or reverse orientation or firefly luciferase (Luc). (B) NIH 3T3 and BHK-21 cells were infected with packaged VEEV replicons expressing PARPrev and PARP12L at an MOI of 20 (inf.u./cell). After incubation for 1 h at 37°C, they were superinfected with packaged VEErep/Luc/GFP replicon at an MOI of 1 (inf.u./cell). Luciferase activity was measured at the indicated time points. (C) BHK-21 and NIH 3T3 cells were infected with packaged VEErep/PARP12L or VEErep/PARPrev replicons at an MOI of 20. Intracellular accumulation of VEEV nsP2 was measured at the indicated time points by Western blotting as described in Materials and Methods. The data were normalized to the level of β-tubulin.