![Fig. 1. Effect of thrombin on JNK1 activation. Platelets were incubated at 37°C with various concentrations of thrombin for 2 minutes (A) or in the presence or absence of thrombin (1 U/mL) for various times (B) as described in Materials and Methods. The reaction was stopped by addition of lysis buffer containing SDS for Western blotting and Triton X-100 for immunoprecipitation. JNK phosphorylation and total JNKs were analyzed by Western blotting using a polyclonal antibody recognizing JNK1-P and JNK2-P and a polyclonal antibody recognizing total JNKs, respectively. For JNK activity, phosphorylated GST-cJun was followed after immunoprecipitation of JNK1, with [32P] ATP. These autoradiographs shown are typical of at least three experiments. (A) Dose-dependent effect of thrombin on JNK activation. Washed platelets were stimulated by incubation with various concentrations of thrombin (0 to 1 U/mL) for 2 minutes with stirring. (B) Time course of thrombin-mediated JNK activation. Washed platelets were stimulated by incubation with 1 U/mL thrombin with stirring.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/94/11/10.1182_blood.v94.11.3800/4/blod42325001w.jpeg?Expires=1723738745&Signature=M3SgSDI-bpJ42YqoZMEC~XlNklwxku2NtiYMCeEYyK3hRUyxKdtrXzODv~aRzrposmSmmb7j2oe3rsKoIrDA9ej-MQ8ufkMsk4r3~54pQkxmt0RmUgSBE5oBccz1k2PjQw4j-BZv~Q6llmq8VNTZZBtDnJiHIh99nef~z8YkGWG0zdYeMdd0P9nC9TONapuYTxnSjbkir4BF9veBrQs8cOHsI4u9eO1rjVx7OLAfTSPgMmWmpIAMiOkRnVsi~-gZspfgYpLYLTBI21WKzMfX5xNRf8D3nQzPJ1nhSCLVyzHxIomnwxaogx~3uK6fmwvbTW7C9FeaV9THymIl~OhvwQ__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Effect of thrombin on JNK1 activation. Platelets were incubated at 37°C with various concentrations of thrombin for 2 minutes (A) or in the presence or absence of thrombin (1 U/mL) for various times (B) as described in Materials and Methods. The reaction was stopped by addition of lysis buffer containing SDS for Western blotting and Triton X-100 for immunoprecipitation. JNK phosphorylation and total JNKs were analyzed by Western blotting using a polyclonal antibody recognizing JNK1-P and JNK2-P and a polyclonal antibody recognizing total JNKs, respectively. For JNK activity, phosphorylated GST-cJun was followed after immunoprecipitation of JNK1, with [32P] ATP. These autoradiographs shown are typical of at least three experiments. (A) Dose-dependent effect of thrombin on JNK activation. Washed platelets were stimulated by incubation with various concentrations of thrombin (0 to 1 U/mL) for 2 minutes with stirring. (B) Time course of thrombin-mediated JNK activation. Washed platelets were stimulated by incubation with 1 U/mL thrombin with stirring.
