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After 2 days of expression, cells were harvested in ice-cold PBS and subsequently lysed in AP-MS buffer (10 mM Tris/Cl pH 7.5, 150 mM NaCl, 10% Glycerol, 0.5 mM EDTA, 0.5% Nonidet P40) supplemented with protease inhibitor cocktail (Roche, Rotkreuz, Switzerland) by sonification, followed by centrifugation (20,000 g for 10 min

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No obstante, la hipertensin no fue un desenlace en los estudios LEAD por lo que no se discrimin el uso de medicamentos antihipertensivos concomitantes