Hippocampal neurons were cotransfected with sybII-pHluorin (1 or 2 μg) and either mCer (1 or 2 μg) or syp-mCer (syp, 1 or 2 μg), and were stimulated with 300 action potentials at 10 Hz. A: Graph displays the mean ΔF/F0 time course for sybII–pHluorin ± SEM normalised to the peak of stimulation. Numbers indicate μg of each construct used. n = 11 except for 2 μg sybII-pHluorin + 2 μg syp (n = 9) and 1 μg sybII-pHluorin + 1 μg mCer (n = 10). p <0.05 (*) against 1 μg sybII-pHluorin + 1 μg mCer, p < 0.05 (#) against 2 μg sybII-pHluorin + 2 μg mCer over time indicated by bar, two-way ANOVA with Tukey’s multiple-comparison post-test. B: One-phase decay curves were fit to the endocytic portion of the traces to determine tau. Graph displays mean tau (s) ± SEM, n as in A, p <0.001 (***) against 1 μg sybII-pHluorin + 1 μg mCer, p < 0.001 (###) against 2 μg sybII-pHluorin + 2 μg mCer, one-way ANOVA with Holm-Šídák’s multiple-comparison post-test.