Figure S2.
Panel A shows two sets of images displaying GCaMP6f fluorescence in astrocytes under resting conditions and after treatment with 1 nanomolar glutamate. The middle section shows calcium two positive amplitude traces, with different colors representing individual traces. The kymograph on the right shows the movement of LAMP1-marked late endosomes or lysosomes over 250 seconds. Panel B is a scatter plot with a mean velocity of late endosomes or lysosomes under 1 nanomolar glutamate treatment, with a p-value of 0.0093 indicated. Panel C shows similar data for astrocytes treated with 1 micromolar glutamate, including GCaMP6f images, calcium two positive amplitude traces, and a kymograph over 150 seconds. Panel D is a scatter plot showing the mean velocity of late endosomes or lysosomes under 1 micromolar glutamate treatment, with a p-value of less than 0.0001. The scatter plots include error bars representing mean standard error of the mean from three independent experiments.
Glutamate pathway stimulation enhances Ca 2+ signaling and modulates lysosomal motility. (A–D) Astrocytes were cotransfected with GCaMP6f and LAMP1-mCherry. Representative trajectories of Ca2+ signals (green) and late endosomes/lysosomes in astrocytes treated with 1 nM (A) and 1 μM (C) glutamate, analyzed using ImageJ TrackMate. Acquired at 0.5-s intervals using Zeiss Airyscan Fast mode. Scale bars: 10 μm. Quantification of the mean velocity of late endosomes/lysosomes under 1 nM (B) and 1 μM (D) glutamate. Data analysis was performed using one-way ANOVA with Dunnett’s multiple comparisons test, **P < 0.01; ****P < 0.0001. Error bars represent the mean ± SEM; n = 3 independent experiments.