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They were washed with and resuspended in PCA medium (5 mM MES pH 5.8, 0.32% Gamborgs B5 basal salt powder with vitamins, 2 mM MgSO 4 .7H 2 O, 3.4 mM CaCl 2 .2H 2 O, 0.342 mM l-glutamine, 58.4 mM sucrose, 444 mM glucose, 8.4 M calcium pantothenate, 4 g/ml biotin, 64.52 g/l ampicillin) at ~300,000 protoplasts per 1.8 ml, before immobilization in 0.75% (w/v) agarose (see below) to acquire confocal microscopy images

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W/S, weakly immobilized/strongly immobilized

135 Suarez-ZamoranoN.FabbianoS.ChevalierC.StojanovicO.ColinD