Construction and replication kinetics of PBj-wt and PBj-Nef202/203GG. (A) Schematic structure of SIV-PBj1.9 genome and PBj-Nef protein. The position of the ITAM (YxxL), SH3-binding motif (PxxPxxP), start of the 3' long terminal repeat (3' LTR) and the D-D-X-X-X-E motif are indicated. (B) or non-stimulated (C) primary macaque PBMCs from 6 animals were infected with the PBj-wt, PBj-Nef202/203GG or PBj-ΔNef virus with an MOI of 1. RT activity was measured in culture supernatants. Error bars, SD. (D) Analysis of RT activity upon infection of non-stimulated primary macaque PBMCs with serial dilutions of PBj-wt or PBj-Nef202/203GG virus. (E) Western blot detection of Nef protein expression in cell lysates of uninfected, PBj-wt-, PBj-Nef202/203GG- and PBj-ΔNef-virus infected C8166 T cells at day 8 p.i. Protein expression of viral Gag, Vpx, Vpr and cellular tubulin was analyzed as control.