Fig. 8.

Analysis of the influence of the duration of centrifugation and pre-centrifugation on the recovery of TF-containing microvesicles. Conditioned media from PAR2-activated MDA-MB-231 cells were centrifuged at 20,000g and 100,000g for intervals ranging from 10 to 120 min. The microvesicles were then resuspended in PBS, and (A) the TF antigen levels and (B) the thrombin generation potential of the recovered microvesicles were measured (n=3, *=p<0.05 vs. the amounts detected in non-centrifuged samples). The conditioned media from AsPC-1, MDA-MB-231 and A375 cells were pre-centrifuged at 10,000g before attempting to recover the microvesicles by centrifuging at 20,000g or 100,000g for 60 min. Similar samples were prepared from microvesicles but without pre-centrifuging. The microvesicles were then resuspended in PBS, and (C) the TF antigen levels and (D) the thrombin generation potential of the recovered microvesicles were measured (n=3, *=p<0.05 vs. the amounts detected in respective samples without pre-centrifuging).