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. Author manuscript; available in PMC: 2011 Aug 15.
Published in final edited form as: Arch Biochem Biophys. 2010 May 15;500(2):181–188. doi: 10.1016/j.abb.2010.05.008

Fig. 1. Tau-degradation in HT22 cells is proteasome, but not ATP-dependent.

Fig. 1

HT22 cells were seeded one day before experiments as described in ‘Materials & Methods’. Cells were incubated in the presence or absence of the proteasome inhibitor, lactacystin (12 μM) for 20h (Panel A). Cells were then lysed and analyzed by immunoblotting using anti-tau and anti-ubiquitin antibodies. One representative out of three experiments is shown. Quantification was performed by using Alpha Ease FC’s “FluorChem 8900”-software. Panel B shows the effect of partial cellular ATP-depletion by KCN and deoxyglucose addition (0.5 mM KCN; 3mM deoxyglucose) on tau levels. The ATP/ADP ratio was calculated from a nucleotide analysis by ion-pair reversed phase HPLC. At the same time point, the content of the 19S proteasomal regulator subunit MSS1, of the tau protein, of GAPDH, and of polyubiquitin conjugates was determined using the corresponding antibodies after immunoblotting. The amounts of human tau and ubiquitin were quantified using AlphaEaseFC’s “FluorChem 8900”-software. Representative results of four independent experiments are shown.