Fig. 1.

CD63/actin positive aggregates in HaCaT cells in the absence and presence of PsVs. HaCaT cells were transfected with CD63-GFP and 1 day later incubated for 3 h at 37 °C without (left panels) or with (right panels) PsVs. Cells were fixed and stained for PsVs and actin with an antibody and fluorescently labeled phalloidin, respectively, and GFP signal was enhanced by nanobodies. Confocal scans from the basal cell membrane were recorded. The linear lookup tables illustrate the channels for CD63-GFP, actin and PsVs in green, red and cyan, respectively. Overlap between green and red is illustrated in yellow and overlap between all three channels in white. Examples of patched structures counted as aggregates in Fig. 2 are marked by arrows in the magnified views. The most left magnified view illustrates patchy CD63-GFP structures that were not counted as aggregates. The two magnified views to the right show aggregates associated with many (left) and few (right) PsVs. For Fig. 2, aggregates were counted per cell base. Large continuous structures, like those often found at the cell periphery (see arrows in overview images) or CD63-vesicular structures in the perinuclear region (see asterisks) were excluded from the analysis