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Figure 5 | Retrovirology

Figure 5

From: Nef-mediated enhancement of cellular activation and human immunodeficiency virus type 1 replication in primary T cells is dependent on association with p21-activated kinase 2

Figure 5

siRNA knockdown of PAK2 in Jurkat cells reduces NFAT activation induced by anti-CD3/CD28 stimulation whether or not Nef is present. A. Jurkat NFAT-Luc cells were transfected with 2 or 10 pmol of PAK2 siRNA or non-targeting siRNA control pool. Three days later, cells were lysed in 1% NP-40 lysis buffer and analyzed by SDS-PAGE/Western blot for PAK2 and β-tubulin expression. B. 200,000 3H cpm of RT activity of the indicated VSV-G pseudotyped virus was incubated overnight with 106 NFAT-Luc cells transfected with 10 pmol of the indicated siRNA. Infected cells were then incubated with 106 α-CD3/CD28 beads for 4 h. Cells were lysed with passive lysis buffer. The lysate was freeze/thawed once and luciferase activity was assayed by luminescence. Average luciferase activity for triplicate samples ± SEM is shown.

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