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. Author manuscript; available in PMC: 2020 May 1.
Published in final edited form as: Thromb Res. 2019 Feb 13;177:59–69. doi: 10.1016/j.thromres.2019.01.017

Fig. 7. PAR4/P2Y12 mediated αIIbβ3 and Rap1 activation and P-selectin expression.

Fig. 7.

PAC-1 binding: platelets from a normal donor were incubated with or without (A) AZD1283 (30 μM) (B), U73122 (10 μM), LY294002 (30 μM), GF109203X (30 μM) and then stimulated with 160 μM AYPGKF for additional 15 min. Platelets were labeled with FITC-Pac1 Ab. P-selectin expression: platelets from a normal donor were incubated with or without AZD1283 (30 μM) (C), U73122 (10 μM), LY294002 (30 μM), GF109203X (30 μM) (D) and then stimulated with 160 μM AYPGKF for additional 10 min. Platelets were labeled with PE-P-Selectin Ab. Representative data are presented from 3 separate experiments. (E) PAR4 mediated Rap1 activation in human platelets from PAR4 variant rs773902 (A/ A homozygote Ala/Ala120 PAR4, T/A heterozygote Thr120/Ala120 PAR4 and T/T homozygote Thr/Thr120 PAR4). Human platelets were stimulated with 160 μM AYPGKF for the indicated times in minutes, and active Rap1-GTP was detected by pull-down assay. Total Rap1 is shown as loading control. Effect of pretreatment with AZD1283 (30 μM) on PAR4 (160 μM AYPGKF) activation on human platelets was determined. Representative data is presented from n = 6, 3 AA, 2 TA and one TT individual in duplicate.