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The attached cells were fixed by immersion into two changes of pre-cooled 20C 200 L 100% (v/v) methanol for 10 min and rehydrated in 500 L 1 PBS (137 mM NaCl, 2.7 mM KCl, 10 mM Na 2 HPO 4 , 1.8 mM KH 2 PO 4 ) at RT for 15 min