tra12161-sup-0001-FigureS1.jpgJPEG image609KFigure S1: Monitoring the trafficking of FLAG-furin from the plasma membrane to the Golgi apparatus in cell populations by FACS. HeLa cells were transfected with FLAG-furin for 24 h. Monolayers were harvested with 5 mm EDTA to obtain a cell suspension and incubated with rabbit anti-FLAG antibodies for 45 min on ice, washed in PBS and either fixed (0 min) or incubated in serum-free media for up to 90 min at 37°C to internalize the antibody-FLAG-furin complexes. Immediately following the incubation, cell suspensions were fixed and permeabilized and stained with Alexa568-conjugated anti-rabbit IgG (red). GM130 was stained with mouse monoclonal anti-GM130 antibodies, followed by Alexa488-conjugated anti-mouse IgG (green). A) Shown are 3D reconstructed confocal images presented as 2D projections. Bar represents 10 µm. B) Histograms of pulse-width values for FLAG-furin and GM130 are shown to provide a complete view of the dataset (n = 2) for each time-point. Thickness of line represents SEM of the dataset at each mean value.

Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.