SDS-PAGE, immunoblotting, and coimmunoprecipitation of lipid raft/caveolae preparations derived from EA.hy926 cells with and without treatment with PC-S195A. (A) After SDS-PAGE of membrane fractions prepared by discontinuous sucrose gradient ultracentrifugation, they were immunoblotted with antibodies directed to PAR-1, EPCR, and caveolin-1. (B) The same as (A) except that the fractions were derived from EA.hy926 cells treated with 50 nmol/L PC-S195A. (C) Total cellular proteins from nontreated and PC-S195A-treated EA.hy926 cells were immunoprecipitated with anti-caveolin-1, anti-PAR-1, and anti-EPCR antibodies, separated on SDS-PAGE and immunoblotted with different pairs of the same antibodies.