Fig. 2

PI3K activation defines a pathogenic element in tau PFF-induced integrin signaling network. (A) A heat map shows the induction in LFC for PI3K signature genes in tau PFF- and OPN-treated primary astrocytes normalized to PBS-treated cells. n = 3 biological repeats. (B) Immunoblotting analysis of phosphorylation (p) of FAK and AKT in primary astrocytes treated with tau PFF or OPN for 2 h. Total (t) FAK and AKT were used as loading controls. Shown are representative blots from three biological repeats. (C) Quantification (mean ± SD) of pAKT and pFAK relative to total tAKT and tFAK (B). ****, p < 0.0001, ****, p < 0.001, ns, not significant by one-way ANOVA. n = 3 biological repeats. (D) Representative gels show time chase of AKT phosphorylation in primary astrocytes treated with tau PFF or SC79 (10 μM). (E) The effect of the indicated inhibitors on AKT phosphorylation in primary astrocytes treated with PBS or tau PFF. The numbers indicate the ratio of pAKT to tAKT. The data represents three biological repeats. (F) iNeurons treated with condition medium from primary astrocytes treated as indicated were stained with a green-fluorescent dye to label viable cells and a red-fluorescent dye to label dead cells. The right panels show enlarged view of the box-indicated areas. (G) Quantification of (F). Error bars indicate mean ± SD. ***, p < 0.001; ****, p < 0.0001 by one-way ANOVA. n = 3 biological repeats