Fig. 2.
Expression and cellular distribution of GlyRα1 wildtype and GlyRα1P1-5A mutant subunits. (A) Western blot analysis of GlyRα1 subunits. 20 μg of membrane preparation was loaded per lane, primary antibody was mAb4a supernatant. Lane 1: α1 wildtype; lane 2: GlyRα1P1-5A; lane 3: protein standard; 48 kDa is indicated. (B) Immunofluorescence: HEK293 cells were transfected on cover slides and treated for immunocytochemistry with mAb4a and goat anti mouse Cy3. For surface expression no Triton X-100 was added. To detect the intracellular protein distribution, the cell membrane was permeabilized with Triton X-100. Controls: untransfected cells or GlyRα1 transfected cells treated with secondary antibody only are shown. Pictures were taken at 400× magnification. Scale bar indicates 10 µm.