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For thin filament determination, NRVM’s were incubated with sterile-filtered Ringer’s relaxation buffer (6 mM potassium phosphate pH 7.0, 100 mM NaCl, 2 mM KCl, 0.1% glucose, 2 mM MgCl 2 , 1 mM EGTA) for 20 min., room temperature, followed by a brief PBS wash to start the WGA staining