FIGURE 7:

Differential effect of GFP-Rab43 on the surface accumulation of G^AE and G. COS7 cells transfected with GFP-Rab43 were subsequently infected with adenoviral vectors expressing (A) G^AE-Ds or (B) G-Ds and grown at the restrictive temperature for 24 h. The cells were then shifted to permissive temperature for 60 min and fixed. Surface G^AE or G was stained in nonpermeabilized cells using the I1 monoclonal antibody, which recognizes an epitope in the ectodomains of both G^AE and G, and an Alexa Fluor 647 secondary antibody. The cells were then imaged by confocal microscopy. The periphery of a cell coexpressing GFP-Rab43 and G^AE-Ds is outlined in A. (C) Quantification of the surface levels of G^AE-Ds and G-Ds in GFP-Rab43– expressing cells shifted to permissive temperature for times ranging from 0 to 120 min (averaged from 12–16 cells in two independent experiments). The difference in the surface delivery of G^AE-Ds at the 120-min time point in the presence and absence of GFP-Rab43 is statistically significant, with p = 0.0003. Scale bars, 5 μm.