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After 24 h transfection, cells were resuspended with ice-cold GST-lysis buffer (20 mM Tris-HCl, pH 8.0, 1% Triton X-100 and 10% glycerol in PBS) supplemented with a protease inhibitor cocktail (Roche) and 50 M Palmostatin B (for SwissKASH-derived samples), and passed 10 times through a 25 G needle