Fig. 3.

Generation of FIV Vif resistant FcaA3Z3. a Representation of FcaA3Z3 protein. Residues investigated are shown. CD cytidine deaminase domain. b, c Several mutants at N-terminal region of FcaA3Z3 were generated. To analyze the sensitivity of FcaA3Z3 mutants to FIV Vif, 293T cells were co-transfected with expression plasmids for FcaA3Z3 wild-type (FcaA3Z3) or indicated mutants and FIV Vif. 48 h later, FcaA3Z3, Vif and tubulin proteins were detected by immunoblot. d Increasing amounts of FIV gag-pol-vif helper plasmid pCPRΔenv that expresses virus-typical levels of FIV Vif without a tag were analyzed for degradation of HsaA3C, FcaA3Z3, FcaA3Z3.A65I and FcaA3Z3.A65I + LI-AA. 293T cells were co-transfected with 150 ng HsaA3C, FcaA3Z3, or A3Z3 mutants and pCPRΔenv (0, 10, 50, 150 or 250 ng). 48 h later, A3 expression was analyzed by immunoblots by using anti-HA antibody. Anti-FIV p24 detected expression of FIV helper plasmid pCPRΔenv, tubulin detection confirmed equal protein loading