FIG.8.

Distribution of GFP, μNS-GFP, and μNS(1-41)-GFP in transfected cells with and without the coexpression of μ2(T1L). (A) CV-1 cells were transfected with 1 μg of pEGFP, pEGFP-M3, orpEGFP-M3(1-41) and 1 μg of pCI-neo as a carrier plasmid per well. Lysates were collected at 18 h p.i. and analyzed by SDS-PAGE and immunoblotting with polyclonal anti-μNS serum (α-μNS) or monoclonal anti-GFP IgG (α-GFP). A band appearing in untransfected CV-1 cells is indicated on the left with an asterisk. The positions of μNS-GFP, μNS(1-41)-GFP, and GFP are indicated on the right. (B) CV-1 cells were transfected with 2 μg of pEGFP (upper left panel), pEGFP-M3 (upper right panel), or pEGFP-M3(1-41) (lower left panel), fixed at 18 h p.t., and immunostained with mouse monoclonal antibody to GFP followed by goat anti-mouse IgG conjugated to Alexa 488. (C) CV-1 cells were cotransfected with 0.25 μg of pEGFP (upper panels), pEGFP-M3 (middle panels), or pEGFP-M3(1-41) (lower panels) and 1.75 μg of pCI-M1(T1L) per well. Cells were fixed at 18 h p.t. and stained for GFP (left column) as described for panel B and μ2 (right column) with rabbit anti-μ2 serum followed by goat anti-rabbit IgG conjugated to Alexa 594. Bars, 10 μm.