Subcellular localisation of EGFP–MyRIP mutants in HUVECs – mutation of the actin-binding region does not perturb endogenous MyoVa recruitment to WPBs. (A) Top, schematic structure of human MyRIP domains and position of the mutated amino acids. Below, confocal immunofluorescence image of a single HUVEC expressing EGFP–MyRIP WT and labelled with phalloidin (red) and specific antibodies against EGFP (green) and VWF (blue). Regions indicated by white boxes are shown as greyscale inserts. (B–D) As for A but for cells expressing EGFP–MyRIP A751P (B), EGFP–MyRIP 4A (C) and EGFP–MyRIP A751P 4A (D). (E,F) Confocal immunofluorescence images of single HUVECs expressing EGFP–MyRIP WT (E) or EGFP–MyRIP 4A (F), and labelled with specific antibodies to EGFP (green) and MyoVa (red). Greyscale images below show, on the same scale, endogenous MyoVa immunoreactivity from regions indicated in the MyRIP-expressing (i) and non-expressing (ii) cells. Scale bars: 10 µm.