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Cells were washed with ice-cold PBS and lysed with a triple detergent lysis buffer supplemented with protease and phosphatase inhibitor cocktail

Plant Physiol 148:16031613 Ohkama-Ohtsu N, Sasaki-Sekimoto Y, Oikawa A, Jikumaru Y, Shinoda S, Inoue E, Kamide Y, Yokoyama T, Hirai MY, Shirasu K, Kamiya Y, Oliver DJ, Saito K (2011) 12-oxo-phytodienoic acid-glutathione conjugate is transported into the vacuole in Arabidopsis