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. 2011 Dec;85(23):12482–12491. doi: 10.1128/JVI.05218-11

Fig. 4.

Fig. 4.

The BC-box motif in FIV Vif is critical for interaction with the Cul5-EB-EC E3 complex. (A) Alignment of partial FIV Vif sequences from 34TF10 with lentiviral Vif and cellular SOCS-box proteins by vector NTI. (B) CrFK cells (106) were cotransfected with 100 ng of VR-fA3Z3-HA, VR-fA3Z2-Z3-HA, or VR-fA3Z2a-HA and 400 ng of VR-FIV Vif-myc or VR-FIV Vif TLQ-myc or VR1012. Stabilities of fA3 proteins were analyzed by Western blotting with anti-HA, anti-myc, and anti-tubulin antibodies. The percentages of fA3s in the presence of FIV Vif TLQ mutant were calculated relative to that in the absence of FIV Vif (set to 100%). 293T cells (C) and CrFK cells (D) (2 × 106) were transfected with 4 μg of VR-FIV Vif-myc or VR-FIV Vif TLQ-myc. Cell lysates were immunoprecipitated with anti-myc antibody, followed by Western blotting with anti-myc, anti-hCul5, anti-hElonginB, or anti-hElonginC antibodies.