FIG 3.

Effects of mutations in the N terminus of GBF1 on interaction with the viral protein 3A. (A) Co-IP of 3A-FLAG with GFP fusions of alanine scanning mutants of GBF1. (B) Co-IP of 3A-FLAG with GFP fusions of GBF1/BIG2 chimeras. Cells were cotransfected with plasmids coding for corresponding GFP-tagged GBF1 mutants and a plasmid coding for 3A-FLAG-Y. IP was performed with anti-FLAG resin. GBF1s were detected with anti-GFP antibodies, and 3A was detected with anti-FLAG antibodies. Actin in the lysates is shown as a loading control. Relative recruitment is calculated by normalizing the GBF1-to-3A signal ratio in the pulldown material of the mutants to that of the positive-control (wt) sample. Each bar is an average of the results from at least three independent experiments. The performance of the corresponding GBF1 constructs in the replication assay is indicated.