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Fig. 4. Insulin granules are differentially regulated by localized Ca2+ upon stimulation. The histograms in A-D were generated similarly to the ones described in Fig. 3. TG, thapsigargin. Measurements were taken 2 minutes after the addition of Bay K8644, nimodipine and glucose, and after 20 minutes of TG treatment. (E) ßTC3 cells were co-transfected with phogrin-EGFP and ECFP-ER, and stimulated with glucose. An image of ECFP-ER was taken prior to acquiring the time-lapse movie of phogrin-EGFP labeled granules. The distance of each granule to the ER, measured by the fluorescence intensity of ECFP-ER at that position, is plotted against the speed of that granule from the tracking analysis. The fast-moving granules, as defined in this paper, are indicated in red. (F) A similar experiment as in E was performed using ECFP-Golgi instead of ECFP-ER. *P<0.001, #P<0.01, &P<0.05, and ^P>0.1, as assessed by paired t-tests. a.u., arbitrary units.





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