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. 2008 May 2;4(5):e1000057. doi: 10.1371/journal.ppat.1000057

Figure 7. Inactivation of the macrophage restriction to SIV by Vpx requires DDB1.

Figure 7

(A) Association of SIV Vpx with endogenous DDB1. Association of DDB1 with wild-type Vpx (VpxWT) and non-ubiquitylated Vpx (VpxM4) was evaluated in 293T cells expressing FLAG-tagged Vpx (left panels) or HA-tagged Vpx (right panels) or IRES-GFP as a control. FLAG and HA immunoprecipitates were immunoblotted with DDB1 or FLAG and HA antibodies (upper panels). Levels of endogenous DDB1 and expressed Vpx in cell lysates were confirmed by Western blotting with a DDB1 antibody and with FLAG/HA antibodies respectively (lower panels). (B) Efficiency of siRNA-mediated silencing of DDB1 expression in COS cells and in macrophages was evaluated by Western blotting with DDB1 antibody at the indicated intervals post siRNA-transfection (ScrΙ-scrambled siRNA control). (C) Impact of DDB1 silencing on SIV and HIV-1 infection of COS cells and macrophages. SIV and HIV-1 infection was gauged from the quantity of viral cDNA (2-LTR) at 24, 48 and 72 h post infection (+, p>0.05; *, p<0.005) (error bars are s.d. of replicate PCRs of a single DNA sample).