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The membranes were blocked in 5% (w/v) nonfat dry milk in phosphate-buffered saline containing 0.1% (v/v) Tween 20 for 1 h at room temperature and then incubated overnight at 4 °C with specific primary antibodies to GCLC, GCLM, multidrug resistance protein 1 (MRP1), or β-actin (Abcam, Eugene, OR) followed by incubation for 1 h at room temperature with a horseradish peroxidase–conjugated secondary antibody (Abcam)