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. 2008 Jul 23;82(19):9657–9667. doi: 10.1128/JVI.00107-08

FIG. 3.

FIG. 3.

Effects of reducing the negative charge within the Nef AC on Nef-mediated PAK2 activation and infectivity enhancement. (A) Nine SF2Nef AC mutants in addition to the wild-type SF2Nef were stably expressed in CEM cells or transiently expressed in 293T cells. The activation of PAK2 by Nef was assayed from extracts of these cells by an in vitro kinase reaction (top). The activity of SF2Nef was set to 100. The negative control (Vector) represents CEM cells transduced with LXSN or 293T cells transfected with pcDNA3.1. The bottom panel is a Western blot analysis of Nef expression in these extracts. (B) A Student's t test was used to compare the PAK2 activation by nine AC mutant Nef proteins. The activation of PAK2 by SF2Nef was set to 100. Note that mutated Nefs are grouped by statistical significance. (C) The SF2 molecular clones, p9B18, p9B18Nef(−), and p9B18 with mutated Nefs were analyzed for their ability to enhance infectivity in a single-round infectivity assay of target HeLa-MAGI cells (n = 6).