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The authors found that benfotiamine enhanced phosphorylation of glycogen synthase kinase (GSK), which leads to the suppression of its activity, and they proposed that this is the mechanism through which benfotiamine exerts its protective effects in the AD model, seeing that GSK has been involved in tau phosphorylation [72] and the production and accumulation of -amyloid in APP overexpressing mice [20]

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Briefly, in vitro translated ATF3 (TnT Quick Coupled Transcription/Translation Systems, L1170) was incubated with E1, E2, and GST-MDM2, ubiquitin with buffer containing 50 mM Tris-HCl, pH 8.0, 50 mM NaCl, 1 mM EDTA, 5% glycerol, 10 mM fresh DTT on ice for 2 h and then add with or without PXR recombinant protein (ProteinOne, Rockville, MD) at 37C for another 2 h