Fig. 2. Glycine induces 36CI– uptake and [Ca2+]i increase in neutrophils pretreated with phorbol myristate acetate (PMA).
(A) 36CI– influx in response to glycine (1 mM) in neutrophils. Neutrophils were incubated with 2 μg/ml PMA for 20 min. Uptake was initiated by adding 36CI– solution and influx was terminated by the addition of 4 ml of ice-cold HEPES butter and rapid filtration. Neutrophils were placed in scintillation vials and solubilized with 0.2 N NaOH for 2 h. EcoLume was added and radioactivity was measured with a Beckman LS-5801 liquid scintillator. (B) Neutrophils were pretreated with 2 μg/ml PMA for 20 min, and glycine (500 μM)-induced reactive oxygen species (ROS) generation was measured. (C) Effect of glycine (1 mM) on [Ca2+]i in vehicle- or PMA-pretreated neutrophils. Neutrophils were pretreated with diphenylene iodonium (DPI) (1 μM) for 15 min before PMA exposure. (A–C) The average of three experiments is shown (mean ± SE). *p < 0.05; **p < 0.01;***p < 0.001 by Student’s t-test. DCF-DA, 2’,7’-dichlorodihydrofluorescein diacetate.
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