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Fig. 1. Cytosolic alkalinization occurs in human eosinophils stimulated with platelet-activating factor (PAF), but not with phorbol myristate acetate (PMA) or IL-5. Purified human eosinophils were loaded with BCECF/AM and stimulated with media control, PAF (3 µM), PMA (2 nM) or IL-5 (0.2 µM) at 37°C. Fluorescence was measured over 90 minutes and was converted to pHi using a standard curve as described in Materials and Methods. Data are shown as mean±s.e.m. (A) Eosinophils stimulated with media control ({circ}; n=17) showed no significant change in pHi compared to pHi at time 0. PAF-stimulated eosinophils ({bullet}; n=17) showed a rapid and sustained cytosolic alkalinization compared to pHi at time 0 that reached a steady-state level approximately 1 hour after stimulation. Initial pHi values for media control and PAF data sets were both 7.1±0.0. (B) Eosinophils stimulated with media control ({square}; n=11), PMA ({blacksquare}; n=9), or IL-5 ({bullet}; n=9) showed no significant changes in pHi compared with pHi values at time 0. Initial pHi values for media control, PMA and IL-5 data sets were 7.1±0.1, 7.1±0.1 and 7.2±0.1, respectively. *P<0.01 (compared to pHi at time 0).





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