FIG. 8.

LCMVSCAT2 minigenome expression is not inhibited by RING finger proteins unrelated to Z. (A) Expression of viral (Vmw110 and IE86) and cellular (PML) RING finger proteins in BHK cells. BHK-21 cells were infected with vTF7.3 (MOI = 3) and transfected with 0.5 μg of RMX-PML (panel i, lane 3), 0.5 μg of pT7-110 (panel ii, lane 4), or 0.5 μg of pGEM-IE86 (panel ii). Protein expression was assayed by Western blotting with either a rabbit anti-PML antibody (panel i, lanes 1 to 3) or a mouse anti-Vmw110 antibody (panel i, lanes 4 to 6) or by immunofluorescence with a mouse anti-IE86 antibody (panel ii). The positions of PML and Vmw110 are indicated on the right, and the molecular size markers are indicated on the left. (B) Inhibition of CAT expression by the LCMVSCAT2 minigenome is not observed with RING finger proteins unrelated to Z. BHK-21 cells were cotransfected with 0.5 μg of pLCMVSCAT2, 1.5 μg of pCITE-NP, 0.1 μg of pGEM-L, 0.1 μg of pTMI-GFP, and increasing amount of pUCIRES-Z, pGEM-IE86, pT7-110, or RMX-PML as indicated at the bottom of the figure. Plasmid pGEM-L was omitted in lane 14. In all of the samples the amount of DNA was kept constant by adding vector pTMI to 2.7 μg. Samples were assayed for CAT activity as described in Materials and Methods. O, origin; Cm, chloramphenicol; MAc, monoacetylated chloramphenicol; DAc, diacetylated chloramphenicol.