Codistribution of CD146 and endothelial cytoskeleton and effect of cytoskeleton-disrupting agents on CD146 distribution.

HUVECs were double stained with anti-CD146 antibody (green) and phalloidin (red) at confluence. (A) CD146 was associated with the actin cytoskeleton. (B) HUVECs double stained for MHC class I (green) and F-actin (red) used as a control. Confluent monolayers of HUVECs were incubated with CCB for 1 hour (C,D) or with EGTA (3 mM) for 5 minutes (E,F). Cells were then double stained with both anti-CD146 antibody (green) and phalloidin (red) (C,E) or with both anti–MHC class I (green) and phalloidin (red) (D,F). Disruption of microfilaments with CCB or EGTA led to a redistribution of CD146 (C,D) compared with findings in untreated cells (A). Magnification × 100.