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. 2013 Nov;347(2):339–345. doi: 10.1124/jpet.113.205104

Fig. 5.

Fig. 5.

PECAM-targeted scFv/TM M388L resistant to oxidative inactivation restores thrombomodulin function of oxidized endothelial cells. (A) Functional resistance to chloramine oxidation of WT versus mutant scFv/TM bound to model PECAM-expressing cells. Model human cell line stably transfected with mouse PECAM control PECAM-negative naive cells (data not shown) were incubated with WT or mutant scFv/TM (left or right bar pairs, respectively) and thrombin alone (dotted line), washed, exposed to 250 µM chloramine-T, incubated further with thrombin and protein C, and the media were examined for APC. Three independent experiments were performed in triplicate, mean ± S.E.M.; *P < 0.05 vs. oxidized scFv/TM-M388L. (B and C) Restoration of TM activity in oxidized endothelial cells. Mouse endothelial MS1 cells were incubated with PBS, WT, or mutant scFv/TM, washed, exposed to chloramine-T (500 or 1000 µM), and the media were examined for APC generated upon addition of thrombin and protein C as in (A). Three independent experiments were performed in triplicate, mean ± S.E.M.; *P < 0.002 for open vs. oxidized scFv/TM-M388L.