Figure 4

Effect of fixation method and antibody choice on co-expression of Vif and APOBEC3G. HeLa cells were transfected with pNL-A1 and pHIV-APO3G as described in figure 3. Cells were grown on cover slips for 24 hr and then either fixed with ice-cold methanol (panels A-C) or with formaldehyde buffer as in figure 3 (panels D-F) and stained with the following combinations of antibodies: (A & D) polyclonal Vif + anti-Myc MAb 9E10; (B & E) anti-Vif MAb #319 + anti-Myc polyclonal antibody; (C & F) anti-Vif MAb #319 + anti-APO3G polyclonal antibody. Vif was visualized using Cy2-conjugated secondary antibodies (green) and APOBEC3G was visualized with Texas red-conjugated antibodies (red). Areas of overlap appear as yellow.