Fig. 5

The CD93-MMRN2 complex is recycled in association with β1 integrin. The CD93-YFP construct was transfected or not into HUVECs. Cells were detached from the plate, resuspended in complete growth medium, plated, fixed at different times after plating, and imaged by immunofluorescence. a, b: Untransfected cells at early and late phases of spreading were stained using phalloidin, anti-CD93 and anti-β1 integrin antibodies. Merged, DIC, and wdc images between CD93 and β1 integrin are shown. In b, the wdc picture shows a high magnification of the CD93^+ve vesicles visible into the dotted square. Scale bars, 6 μm (a), 4 μm (b). c, d: Transfected early and late spreading ECs were stained for β1 integrin. Exogenous CD93 was imaged as green. Overlay of stained cells and wdc images are shown. Scale bars, 8 μm (c) and 7 μm (d). e, f: Immunofluorescent staining of CD93 and active β1 integrin (12G10) in untransfected cells at early and late phases of attachment to the substrate. Merged and wdc images are shown. In f, dotted lines indicate cell boundary. Scale bars, 8 μm (e) and 13 μm (f). For each condition, Manders and Costes quantitative analyses of β1 integrin colocalization with CD93 and vice versa are reported (n = 4–8 cells)