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. 2006 Aug;80(15):7535–7545. doi: 10.1128/JVI.02741-05

FIG. 4.

FIG. 4.

Apoptin localization signals are modular and can cooperate in trans to confer cell type-specific localization. (A) H1299 (left panels) and PFF (right panels) cells expressing GFP-Ap-pmNLS, dsRed-Ap-pmNES, or both were monitored by fluorescence microscopy. (Far right) Quantification of the nuclear/cytoplasmic GFP signal ratio for the images presented in the panels on the left. (B) H1299 cells were transfected with GFP-Apoptin mutants and 12 h later infected with Ad-Apwt. Apoptin multimerization was monitored by immunoprecipitation of Flag-tagged Apoptin using an α-Flag affinity resin, followed by analysis of the immunoprecipitate (IP) for GFP-Apoptin by immunoblotting with an α-GFP antibody. Expression of the GFP-Apoptin mutants and Flag-Apoptin were also monitored in whole-cell extracts (WCE). Samples were normalized by equivalent cell number. (C) H1299 (left panels) and PFF (right panels) cells were cotransfected with GFP-Ap(1-88) and either dsRed-Apwt (top), dsRed-Ap-pmNES (middle), or dsRed-Ap(82-121) (bottom) and monitored by fluorescence microscopy. (Far right) Quantification of the nuclear/cytoplasmic GFP signal ratio for the images presented in panels on the left.