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. Author manuscript; available in PMC: 2010 Jan 24.
Published in final edited form as: Mol Ther. 2008 Jun 17;16(8):1450–1458. doi: 10.1038/mt.2008.127

Figure 5. Mechanism of endocytosis of anti-ICAM carriers of various geometries.

Figure 5

(a) Fluorescence microscopy showing formation of actin stress fibers (stained by red Alexa Fluor 594 phalloidin) upon incubation of activated HUVECs with fluorescein isothiocyanate–labeled anti-ICAM/spheres (0.5 and 5 μm diameter) or anti-ICAM/disks (0.1 × 1 × 3 μm) for the indicated time. Particles in the cell surface look blue due to counter-staining with blue Alexa Fluor 350 goat anti-mouse immunoglobulin G. Scale bar = 10 μm. (b) Internalization (1 hour) of anti-ICAM spherical particles of various sizes (0.1, 1, and 5 μm diameter) and elliptical disks (0.1 × 1 × 3 μm) in the presence of two pharmacological inhibitors of actin filaments (0.5 μmol/l cytochalasin D or CytD, and 0.1 μmol/l latrunculin A or LatA). (c) The effects of pharmacological inhibitors of clathrin-coated pits (50 μmol/l monodansyl cadaverine, MDC), caveolar-mediated endocytosis (1 μg/ml filipin, Fil), a common inhibitor to macropinocytosis and CAM endocytosis (3 mmol/l amiloride, Amil), and a macropinocytosis inhibitor (0.5 μmol/l Wortmannin, Wort), were tested as in (b). Data are mean ± SEM (n ≥ 25 cells, two experiments). Calculated with respect to control cells (%Ct). ICAM, intercellular adhesion molecule 1.