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Figure 6


Fig. 6. Association of tau protein with murine {gamma}-satellite DNA. Electrophoretic mobility shift assays (EMSA) were performed to analyze the interaction of purified tau protein with murine {gamma}-satellite DNA sequences. (A) Different concentrations of purified tau protein (100 ng, 200 ng, 300 ng and 400 ng) were incubated with the 32P-labeled {gamma}-satellite DNA probe of 936 bp in the presence of 0.5 µg ssDNA as random non-sequence specific competitor DNA. The gels were dried and the radioactivity was visualized using PhosphorImager and the computer program ImageQuant. (B) Indicated amounts of tau protein or HMGI protein were incubated with the {gamma}-satellite DNA probe, either labeled (lanes 1-5) or unlabeled (lanes 6-13) in the presence of 0.5 µg ssDNA unlabeled competitor. No satellite DNA was added in lanes 9 and 13. The complexes were resolved in a non-denaturing TBE-polyacrylamide gel. The gel was either, dried and autoradiographed (lanes 1-5) or transferred to a nitrocellulose membrane and immunoblotted using either the Tau-5 monoclonal antibody (lanes 6-9) or an anti-HMGI polyclonal antibody (lanes 10-13). The arrows indicate the shift observed for the complexes owing to the interaction with tau and the free-labeled probe.





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