American Society of Hematology

Fig. 8.

$Fig. 8. Movement of PKC-phosphorylated adducin to cytosol with thrombin activation. / Column-purified platelets were activated with dilute thrombin (0.1 U/mL) for increasing times and fractionated by high-speed centrifugation (126 000g for 3 hours) into supernatant and pellet fractions. Samples were run on SDS-polyacrylamide gel electrophoresis and Western blotted; relative optical density of bands was quantitated by using NIH Image software. (A) Typical experiment showing increasing adducin in supernatant versus decreasing adducin in pellet. (B) Distribution of PKC-phosphorylated adducin between supernatant and pellet fractions over time (n = 2). (C) Platelets were preincubated with calpeptin (200 μg/mL for 30 minutes), stimulated with thrombin (1 U/mL), then fractionated as above. PKC-phosphorylated adducin continued to increase in the supernatant while decreasing in the pellet fraction.$

Movement of PKC-phosphorylated adducin to cytosol with thrombin activation.

Column-purified platelets were activated with dilute thrombin (0.1 U/mL) for increasing times and fractionated by high-speed centrifugation (126 000g for 3 hours) into supernatant and pellet fractions. Samples were run on SDS-polyacrylamide gel electrophoresis and Western blotted; relative optical density of bands was quantitated by using NIH Image software. (A) Typical experiment showing increasing adducin in supernatant versus decreasing adducin in pellet. (B) Distribution of PKC-phosphorylated adducin between supernatant and pellet fractions over time (n = 2). (C) Platelets were preincubated with calpeptin (200 μg/mL for 30 minutes), stimulated with thrombin (1 U/mL), then fractionated as above. PKC-phosphorylated adducin continued to increase in the supernatant while decreasing in the pellet fraction.

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