Figure EV2. Aβ_1–42 oligomers increase Ca²⁺ levels in distal neurites in an NMDA‐dependent manner.

1. Hippocampal neurons were grown at low density (5,000 cells cm⁻²) for > 10 DIV and loaded with Fura2‐AM. Ca²⁺ levels were measured before and during perfusion with Aβ_1–42 oligomers in the presence or absence of the NMDA receptor antagonist MK801 and the Ca²⁺‐chelator BAPTA‐AM. Relative changes in Fura2‐AM fluorescence intensity were plotted as means of 35–42 neurites imaged in two independent experiments.
2. Cumulative Ca²⁺ levels represented by the area under the curves in (A). Mean of 27–42 neurites imaged in two independent experiments. ***P < 0.001; one‐way ANOVA with Bonferroni's multiple comparisons test. ^### P < 0.001, t‐test control Aβ_1–42 vs. DMSO.
3. Micrographs of representative axonal segments quantified in (A and B). Fura2‐AM levels before (two most left panels, grayscale and pseudocolor image) and after (right panel, pseudocolor image) Aβ_1–42 perfusion at the indicated experimental conditions are shown. Scale bar, 25 μm.