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. 2012 May;86(9):4856–4867. doi: 10.1128/JVI.06329-11

Fig 1.

Fig 1

Association of sdAb19 and Neffins with Nef. (A) Schematic representation of Neffins resulting from the fusion of sdAb19 to RRT-SH3 B6 or C1 domain (highlighted) using a flexible GGSGG loop. TAG, c-Myc-epitope tag. (B) Intracellular distribution of Nef-GFP and sdAb19, Neffin B6, or C1. HeLa cells were transfected with the plasmid for the expression of the c-Myc-tagged sdAb19 or Neffins in combination with the plasmid for the expression of Nef-GFP. SdAb19 and Neffins were detected by indirect immunofluorescence with anti-c-Myc. The colocalization of Nef-GFP with sdAb19 and Neffins B6 and C1 is shown in white (right panels), and the Pearson's coefficient (r) for the evaluation of the level of colocalization is indicated on the right. (C) Coimmunoprecipitation of Nef-HA with sdAb19 or Neffins. 293T cells were transfected with the plasmid for the expression of the c-Myc-tagged sdAb19 or Neffins in combination with the plasmid for the expression of either Nef-HA or HA (mock). Control corresponds to cells that did not express sdAb19 or Neffins. Cell lysates (right images) were submitted to immunoprecipitation with anti-c-Myc (left images). Immunoprecipitates were analyzed by Western blotting (WB) using anti-c-Myc (bottom) or anti-HA (top). IP, immunoprecipitation.