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Fig. 7. (A) Ionomycin releases Ca²⁺ from the ER but not from acidic stores. (a) NAADP-induced additional Ca²⁺ response from the secretory granule area of the permeabilized cell. (b) Averaged traces from the last 100 seconds (dotted boxes) of the experiments shown in a and c (from the granular (blue) and basal (red) areas, respectively) with application of 100 nM NAADP in the continuous presence of 10 µM ionomycin (n=6, P<0.005, asterisk shows time point when the amplitudes at the granular and basal areas have been compared using t-test). Bars represent standard errors. (c) No response is seen in the basal area. (B) IP₃-induced additional Ca²⁺ response in the presence of 10 µM ionomycin from the secretory granule area of a permeabilized cell. (C) Nigericin-induced additional Ca²⁺ response in the presence of 10 µM ionomycin from the secretory granule area of a permeabilized cell. In A, B and C cells were loaded with Fluo-5N AM. (D-E) Thapsigargin does not discharge acidic Ca²⁺ store in intact cells. (D) Subsequent applications of 10 nM and then 10 µM thapsigargin in the absence of external Ca²⁺ did not prevent an addition small CCK-induced (1 nM CCK) Ca²⁺ release. Cells were loaded with Fluo-4 AM. (E) Averaged traces from 100 seconds of the experiment shown in D (dotted blue box) shows in detail the response to a subsequent application of 1 nM CCK in the absence of external Ca²⁺ (n=8, P<0.0005; asterisks show time points at which relative fluorescence was compared using a t-test). Bars represent standard errors.

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