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Fig. 2. Differential effects of ionic strength on the Slac2-a·myosin-Va complex according to whether exon F is present. COS-7 cell lysates containing FLAG-myosin-Va-GT (or FLAG-myosin-Va-F-GT) (A) or containing FLAG-brain myosin-Va-tail (or FLAG-MC myosin-Va-tail) (B) were incubated with T7-Slac2-a beads in the presence of the concentrations of NaCl indicated. Proteins that bound to the beads were first analyzed with HRP-conjugated anti-FLAG tag antibody (Blot: anti-FLAG, top panels), and then with HRP-conjugated anti-T7 tag antibody (Blot: anti-T7, bottom panels). Note that the binding of the MC-type myosin-Va to Slac2-a was resistant to ionic strength (up to 750 mM NaCl) (lanes 5-8), whereas binding of brain myosin-Va to Slac2-a was highly sensitive to ionic strength (lanes 1-4). (C) Tail constructs of brain-type and MC-type myosin-Va used in this study.





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