Fig. 5

MIF regulation of tau phosphorylation through activated astrocytes. a Primary cultures of astrocytes (Ast) from WT and Mif ^−/− newborn mice were treated with DMEM with or without high glucose (75 mM and 150 mM) for 12, 24, 48, and 72 h. The cell lysate was analyzed by Western blotting for the expression of GFAP. The data are presented as densitometric values after normalization against the GAPDH level and are shown as the means ± SEM from at least three separate experiments; *p < 0.05, **p < 0.01 scompared with cells without stimulation. For b and c, freshly isolated neurons from WT and Mif ^−/− mice were cultured for 48 h with conditional medium (CM) of WT (b) and Mif ^−/− (c) astrocytes that were exposed to high glucose (75 mM and 150 mM) or normal medium (c, DMEM) for 24 and 48 h. Western blotting was performed to detect phosphorylated tau (pT205) in primary neurons. The blots were quantified by densitometry after normalization against total tau (Tau5). All data shown are means ± SEM of multiple experiments (n ≥ 3), each in dulplicate. c Control. * and ^# p < 0.05, ** and ^## p < 0.01 compared with medium without high glucose