Fig. 6
p47phox in astrocytes affects tau phosphorylation by activating astrocytes. Freshly isolated neurons from WT mice were cultured for 48 h with conditioned medium (CM) of WT and Ncf1−/− astrocytes that were exposed to high glucose (75 mM and 150 mM) or normal medium for 24 h (a) and 48 h (b). Representative Western blots showing tau hyperphosphorylation at S199, T205, S396, and S404 in primary neurons. The levels of total tau (Tau5) were also measured. Quantification of the immunoreactivity of Western blots, normalized against total tau (Tau5). One-way ANOVA: a, WT-Ast, pS199 F (2, 32) = 7.126 p = 0.0028, pT205 F (2, 8) = 5.410 p = 0.0327, pS396 F (2, 19) = 11.07 p = 0.0006, pS404 F (2, 21) = 3.466 p = 0.0500; KO-Ast, pS199 F (2, 9) = 0.2762 p = 0.7649, pT205 F (2, 9) = 2.046 p = 0.1851, pS396 F (2, 9) = 2.323 p = 0.1537, pS404 F (2, 9) = 0.3956 p = 0.6844; b, WT-Ast, pS199 F (2, 9) = 10.95 p = 0.0039, pT205 F (2, 8) = 7.484 p = 0.0147, pS396 F (2, 8) = 14.10 p = 0.0024, pS404 F (2, 8) = 6.255 p = 0.0231; KO-Ast, pS199 F (2, 16) = 0.5406 p = 0.5927, pT205 F (2, 16) = 1.599 p = 0.2327, pS396 F (2, 16) = 1.380 p = 0.2800, pS404 F (2, 16) = 2.149 p = 0.1490. Data are mean ± SEM from three separate experiments, each in duplicate or triplicate. *p < 0.05, **p < 0.01, ***p < 0.001 compared with medium without high glucose