Overexpression of dominant negative Rab5A variant inhibits the endocytosis of FVIIa-EPCR in CHO-EPCR cells.

<p>CHO-EPCR cells transduced to express dominant negative Rab5A mutant were incubated with AF488-FVIIa (50 nM) for 1 h at 4°C, and internalization was induced at 37°C as described in Fig. 3. The cells were immunostained for EPCR and Rab5. They were then analyzed for immunofluorescence of EPCR and Rab, and fluorescence of AF488-FVIIa. The 5^th and 6^th panels are the digitally enlarged images of the insets of the 2^nd and 4^th panels, respectively. The extreme right panel was the same panel that was shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0059304#pone-0059304-g003" target="_blank">Fig. 3</a>, but re-inserted here to illustrate differences in AF488-FVIIa and EPCR accumulation in the REC between control and Rab5A DN expressing cells. Therrow indicates the accumulation of AF488-FVIIa at REC.