![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=1735847751&Signature=Ai4R5dfhL4-d-ZpVWqHnzO22HYgyFqQa7XIFBJ2l3YmcITdvWSmdk3HtWzZo1bYC1TQZeKV3vD~iG49I4GDdWmne75adDPxlV7hwFeP1-54ZUzzWQ-IjUKrDLAvKriUhihzMQL9oiTdByPKne6yWb7AHh6WN17KNWG1Ud1ifodpCjhzx~qsEaPJwRi5PwZGEM1M-8RcO~nFX3XMNnQB0h2iMiwByaZW5uSLemRVcDRNLQuMhQm2BcBGItviuhTHf7nwGZv5nTMRF4M0THwP-6DC5LMxZahbv~vh4PcUX~ZbBGoCI3gV4W8vYEjTrNDkoQZZ-SAEoUomldDu1OdVCUw__&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.
