Fig. 6

Inhibition of MIF during high glucose treatment of primary astrocytes abrogated the ability of the CM to stimulate tau phosphorylation in neurons. Primary astrocytes from WT mice were treated with 75 mM (a) or 150 mM (b) of high glucose in the presence or absence of the MIF inhibitor ISO-1 (50 μM and 100 μM). CM collected after 24 h and 48 h was added to primary cultures of mouse neurons. After 48 h, the neurons were harvested and the level of tau phosphorylation (pT205) was determined by Western blotting. Densitometry readings of the blots were normalization against those of total tau (Tau5). The level of tau phosphorylation in the untreated samples (DMEM without high glucose or ISO-1; marked as C in the first lane) was set at 1.0, with which other samples were compared. Data presented in bar charts are means ± SEM of three individual experiments. *p < 0.05, **p < 0.01 vs. high glucose medium without ISO-1