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Fig. 8 | Journal of Neuroinflammation

Fig. 8

From: Role of transient receptor potential ankyrin 1 channels in Alzheimer’s disease

Fig. 8

PP2B activation is vital in Aβ-induced inflammation in astrocytes. (a) Immunostaining of hippocampus specimens from 8-month-old APP/PS1 Tg and APP/PS1 Tg/TRPA1−/− mice with anti-GFAP and anti-PP2B antibodies, then FITC- or Texas red-conjugated secondary antibody. Bar = 50 μm. GFAP-positive cells (red color) denote astrocytes and FITC-positive cells (green color) denote PP2B protein. (b and c) Western blot analysis of PP2B and its activity in brain lysates from 8-month-old WT, TRPA1−/−, APP/PS1 Tg and APP/PS1 Tg/TRPA1−/− mice. (d) Primary astrocytes from WT and TRPA1−/− mice were treated with Aβ (2 μM) for 30 min. (e) WT astrocytes were pretreated with HC (10 μM), 0.5 mM EGTA or 0.5 mM EDTA for 2 h, then Aβ (2 μM) for 30 min. (f-k) WT astrocytes were pretreated with cyclosporin A (CsA, 100 nM) or fenvalerate (Fen, 100 nM) for 2 h, then with Aβ (2 μM) for 24 h (f-i) or 1 h (j and k). ELISA of (f-i) IL-1β, IL-6, IL-4 and IL-10 in culture medium and (j, k) NF-κB and NFAT DNA binding activity in cell lysates. (l) Immunostaining of astrocytes from treated WT mice with anti-GFAP antibody, then FITC-conjugated secondary antibody. Bar = 100 μm. Data are mean ± SEM from 5 independent in vitro experiments or from 8 mice in each group. *, P < 0.05 vs. vehicle or WT mice. #, P < 0.05 vs. Aβ treatment or APP/PS1 Tg mice

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