Figure 5.
Expression of Nef from HIV-1SF2 induces a perinuclear accumulation of human CCR1, CCR2, CCR3, and CCR5. (A and C) Stable CHO transfectants or (B) human TZM hCCR5 high cells were plated on coverslips and transfected with expression constructs encoding for Nef.GFP or GFP alone (A), Nef.GFP or NefAxxA.GFP (B), or Nef.RFP or RFP alone together with Rab11.GFP (C). One day after transfection, receptors were indirectly immunostained with Alexa Fluor 568 (red) (A) or with Alexa Fluor 660 (blue) (B and C) together with indirect immunostaining of TGN46 with Alexa Fluor-568 (red) (B). Images were taken with a Zeiss LSM510 confocal microscope with a 100× PLAN-APO objective. Shown are individual sections (GFP-, RFP-, Alexa Fluor 568- or Alexa Fluor 660-stained receptors, or Alexa Fluor 568-stained TGN46; and merged images) from the center of representative cells. White asterisk indicates cells with perinuclear CKR accumulation. White bars in merged images, 10 μm.