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Figure 4 | Retrovirology

Figure 4

From: Self-association of the Lentivirus protein, Nef

Figure 4

SIV MAC239 Nef oligomerizes and also forms heterologous oligomers with SF2Nef. (A) 293T cells were transfected to express tagged SIVMAC239-HFNef and untagged SIVMAC239Nef, singly and in combination. Membrane protein fractions were prepared and Nef expression was determined by Western blot analysis (Input). Immunoprecipitations were performed with anti-HA monoclonal antibody and the immunoprecipitates analyzed by SDS/PAGE followed by Western blotting with monoclonal anti-SIV antibody (IP:α-HA). Arrows, total tagged Nef (T) and untagged Nef bound to tagged Nef (U). (B and C) 293T cells were transfected to express SF2Nef and SIVMAC239Nef singly and in combination. Membrane (B) and soluble (C) protein fractions were prepared and Nef expression determined by Western blot analysis with anti-HIVNef monoclonal antibody (Input, upper panels) and anti-SIVNef monoclonal antibody (Input, lower panels). The membrane and soluble fractions were also immunoprecipitated with polyclonal anti-HIV-1Nef (IP: α-HIV-1Nef) and polyclonal anti-SIVNef (IP: α-SIV Nef). The immunoprecipitates were analyzed by Western blot with monoclonal anti-HIV Nef (upper panels) and monoclonal anti-SIVNef (lower panels).

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