Aggregation induced by 4N1-1 is unaffected in IAP-deficient mice and in platelets treated with anti-IAP antibodies.
(A) Human platelets were pretreated for 10 minutes with 50 μg/mL F(ab′)2 fragment of antihuman IAP monoclonal antibody B6H12. (B) Mouse platelets were pretreated for 10 minutes with 50 μg/mL antimouse IAP antibodies mAb 301 and mAb 430. Platelets were then stimulated with 25 μM 4N1-1 or 50 μM 4N1K. (C) Mouse platelets from WT mice and IAP-deficient mice (IAP−/−) were stimulated with 25 μM 4N1-1. Aggregation was monitored by light transmission. Arrows indicate addition of agonist. (D) Mouse platelets from WT and IAP-deficient mice were stimulated with 25 μM 4N1-1. Whole-cell lysates were resolved by 12.5% SDS-PAGE and analyzed with Western blotting using an antiphosphotyrosine antibody. Results are representative of experiments using samples from 3 different donors and 2 different mice.
