Fig. 4. BAD-LAMP is present in vesicles clustering in specific areas of the neurons. Immunofluorescence confocal microscopy of cortical neurons. (A) Staining for BAD-LAMP (red) and cholera toxin (GM1, green). (B) Staining for BAD-LAMP (red) and PrP (green). (C) Staining for BAD-LAMP (red) and N-CAM (green). BAD-LAMP is expressed in small vesicles clustered in neurites and accumulates in areas lacking surface semi-ordered lipid microdomain resident proteins (arrowheads). (D) THY1 labelling (green) defines the zones in which BAD-LAMP vesicles accumulate (red). However, THY1 (red) is present at the cell surface and does not co-localize with BAD-LAMP as seen at higher magnification (bottom panels, arrowheads). (E) Cholesterol depletion disrupts cluster organization and induces BAD-LAMP (red) and cholera toxin (GM1, green) co-localization. Bars, 20 µm; 10 µm for THY1 high magnification.