Author response image 1. a and b – Cumulative histograms of single vesicle fusion event amplitudes at room temperature (24 °C) and physiological temperature (34 °C), measured with vGluT1-pHluorin.

c and d – Plot of decay time constant as a function of dwell time duration at room temperature (24 °C) and physiological temperature (34 °C), measured with vGluT1-pHluorin. Solid black lines: linear fit, showing no correlation (R² ~ 0). e – Cumulative distribution of decay time constants for single vesicle fusion events showing no differences at the two measured temperatures. Inset: mean value ± SEM. N of experiments are indicated in Figure 3 legend of the main manuscript.