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. Author manuscript; available in PMC: 2016 Jun 1.
Published in final edited form as: J Thromb Haemost. 2015 Apr 23;13(6):1064–1072. doi: 10.1111/jth.12901

Fig. 2.

Fig. 2

Bilirubin inhibits the proteolytic cleavage of plasma VWF by ADAMTS13. A. Purified plasma VWF (50 nM) was incubated with (+) or without (−) recombinant ADAMTS13 (rA13) (10 nM) in the absence (−) or presence (+) EDTA (20 mM) or increasing concentrations of unconjugated bilirubin (0-5 mM). The proteolytic cleavage of VWF was determined by Western blotting after being separated with 1% SDS-agarose (top) or 5% SDS-polyacrylamide (bottom) gel electrophoresis. UL and P indicate the uncleaved VWF multimers and the cleavage product (~350 kDa), respectively. B. The relative amount of cleavage product of VWF in A-bottom was quantified by densitometry using ImageJ and plotted against the concentrations of unconjugated bilirubin. The data indicate the means ± standard errors (SE) from three independent experiments (n=3).