Acquisition of MyRIP expression is coincident with more peripheral WPBs. (A) HUVECs were transfected with VWF-GFP, and cells were fixed and stained for immunofluorescence after 4, 5, 6, 7, and 8 hours. Maximum intensity projection of confocal images was constructed, and expression of MyRIP (red), total VWF (blue), and newly made VWF-GFP was determined (images are shown at 4, 6, and 7 hours). Scale bar represents 10 μm. The distance of VWF-GFP–positive WPBs from the nucleus was determined from 5 cells at each time point and the MyRIP-negative (blue) and MyRIP-positive (red) WPB plotted. Black bars represent the mean distance values; red error bars, SD. (B) Images taken from a 5-minute movie showing the acquisition of Rab27a-mcherry and MyRIP-GFP. Time points are indicated. Scale bar represents 5 μm. Images were acquired on a SP5 confocal microscope with a 2.513A pinhole at one frame every 5 seconds.