Figure 7

R105 and R106 are critical for Nef oligomerization. (A), 293T cells were transfected to express SF2-HFNef (WT), SF2-HFNefR105D (R105D), SF2-HFNefD123R (D123R), and SF2-HFNefR105D/D123R (R105D/D123R). The presence of Nef dimers was detected by treatment of intact cells with 0.5% formaldehyde (+). No formaldehyde (-). (B) Transduced CEM cells expressing SF2Nef, SF2NefR105A, SF2NefD123E, SF2NefR105D, SF2NefD123R, or SF2NefR105D/D123R plus the vector (LXSN) positive control were assayed for CD4 and MHCI cell surface expression by flow cytometry analysis (left) , PAK2 activation (middle), and level of Nef expression by Western blot analysis (right). (C) Role of R105 and R106 in dimerization was investigated as in (A). SF2-HFNefR105A (R105A), SF2-HFNefR106A (R106A).