Characterization of mGPVI-Fc. (A) Three micrograms purified mGPVI-Fc was subjected to SDS-PAGE under nonreducing (NR) and reducing (R) conditions, and the gel was stained with Coomassie brilliant blue. (B) Mictrotiter plates were coated with mGPVI-Fc and then incubated with the indicated HRP-labeled antibodies followed by TMB development (JAQ1, 2, and 3 [anti-GPVI], p0p4 [anti-GPIbα]). (C) Dose-dependent binding of GPVI-Fc to immobilized collagen (50 μg/mL) and CRP (2 μg/mL) was measured by ELISA (mean ± SD of 3 experiments). (D) mGPVI-Fc partly inhibits collagen- and CRP-induced mouse platelet aggregation. Under stirring conditions, mouse PRP was preincubated with 20 μg/mL GPVI-Fc and then stimulated with the indicated concentrations of collagen or CRP. Light transmission was recorded on a Fibrintimer 4-channel aggregometer. Results shown are representative of 6 experiments.