Supporting Information

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Movie S1: Total internal reflection fluorescence microscopy (TIRFM) movie of insulin-EGFP-labeled granules in MIN6 cells. The excitation wavelength was 491 nm (Cobolt Calypso), the objective was a Zeiss α-Plan-Fluar (100×, 1.45 NA), the angle of incidence was 68° and the calculated decay constant of the evanescent field was 84 nm. The acquisition rate was 8.3 Hz, exposure time 50 milliseconds. The cells were continuously perifused at a rate of 0.2 mL/min, the temperature was 37.0 ± 0.1°C. Three minutes prior to the acquisition of this sequence, the K+ concentration in the Krebs-Ringer perifusion medium was increased by switching to a medium containing 40 mM K+. The lag time before the perifusion chamber was reached by the medium was 1 min. Multiple transient brightenings indicate exocytotic activity.

Movie S2: Same experiment and same acquisition conditions as shown in Movie S1. In addition to 40 mM K+, 10 µM nifedipine was present in the perifusion medium. The exposure time of the MIN6 cells to nifedipine prior to the acquisition of the sequence was 5 min.

TRA_1231_sm_SuppMovie1.mov835KSupporting info item
TRA_1231_sm_SuppMovie2.mov612KSupporting info item

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