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. 2006 Dec 20;81(5):2391–2400. doi: 10.1128/JVI.01601-06

FIG. 4.

FIG. 4.

Effect of mutations within the zinc finger motifs of ZAP on its antiviral activity. (A) Rat2 cells expressing wild-type NZAP-Zeo or mutants containing single amino acid exchanges (H86K, C88R, C168R, or H191R) were infected with Zaire-EBOV, Sudan-EBOV, or MARV Musoke at an MOI of 0.01. Seven days p.i., the infectious virus titer was determined by immunofocus assay. Data represent means and ranges of duplicate infection experiments. Dashed line, immunofocus assay detection limit. (B) Level of NZAP-Zeo mutant protein expressed by cell lines as detected by Western blot analysis. Rat2-Zeo cells, Rat2-NZAP-Zeo cells, and Rat2 cells expressing NZAP-Zeo mutants containing the single amino acid exchange H86K, C88R, C168R, or H191R were lysed in Laemmli loading buffer. Proteins were separated by SDS-PAGE and transferred onto a membrane. NZAP-Zeo and NZAP-Zeo mutants were detected using a polyclonal anti-ZAP antibody. As a loading control, detection of GAPDH using an anti-GAPDH monoclonal antibody was performed.