FIG. 5.

Alteration of the distribution of PML NB components changes expression from targeted transgene. (A) A total of 100 ng biotinylated CMV-pGL3 (lanes 1 to 3 and 7 to 9) or unbiotinylated CMV-pGL3 (lanes 4 to 6 and 10 to 12) was cotransfected with BBP-PMLIV to achieve targeting of the biotinylated vector but not the unbiotinylated control. Cells were also cotransfected with increasing amounts of PMLVI (lanes 1 to 6) or PMLIV (lanes 7 to 12) as indicated. Data are shown normalized to that of Renilla luciferase. (B) A total of 100 ng SV40-pGL3-TetO was cotransfected with 100 ng TetR-PMLIV (white bars) or PMLIV as a nontargeted control (black bars). Increasing amounts of Sp100 were cotransfected with the targeting constructs as indicated. Luciferase activity was normalized relative to the well containing no Sp100. (C) A total of 100 ng biotinylated (white bars) or unbiotinylated (black bars) CMV-pGL3 was cotransfected with 100 ng BBP-PMLIV. Increasing amounts of Sp100 were cotransfected with the targeting constructs as indicated. Luciferase activity was normalized relative to the well containing no Sp100. (D) A total of 100 ng SV40-pGL3-TetO was cotransfected with 100 ng TetR-PMLIV (white bars) or PMLIV as a nontargeted control (black bars). Increasing amounts of 3K-PMLIV were cotransfected with the targeting constructs as indicated. Luciferase activity was normalized relative to the well containing no 3K-PMLIV. (E) A total of 100 ng biotinylated (black bars) or unbiotinylated (white bars) CMV-pGL3 was cotransfected with 100 ng BBP-PMLIV. Increasing amounts of 3K-PMLIV were cotransfected with the targeting constructs as indicated. Luciferase activity was normalized relative to the well containing no 3K-PMLIV. *, P was <0.05; **, P was <0.01. Error bars represent standard deviations of three separate measurements. −, absence of; +, presence of.