Fig. 5

Aβ elicits TRPA1-dependent Ca²⁺ influx in astrocytes. (a) Immunostaining of primary astrocytes from WT mice with anti-IgG and anti-TRPA1 antibody, then FITC-conjugated secondary antibody. (b) Western blot analysis of TRPA1 and α-tubulin protein levels in astrocytes treated with 2 μM Aβ for the indicated times (0–18 h). (c) Ca²⁺ influx in astrocytes treated with 2 μM Aβ for 0–60 min. Fluo-8 calcium assay of intracellular level of Ca²⁺ in (d, e) primary astrocytes from WT and TRPA1^−/− mice treated with Aβ (2 μM) for 2 min and (f, g) astrocytes pretreated with HC030031 (HC) 10 μM, EGTA 0.5 mM and EDTA 0.5 mM for 2 h, then treated with Aβ for 2 min. Fluorescence images were photographed by fluorescence microscopy. Bar = 100 μm. Data are mean ± SEM from 5 independent experiments. *, P < 0.05 vs. vehicle or 0 min. #, P < 0.05 vs. WT Aβ treatment or vehicle