Brains, spinal cords, and livers were isolated, desheathed in saline, and immediately frozen on dry ice. Lysates were prepared by homogenizing the tissues in T-PER buffer (Thermo Fisher; 78510) with a protease inhibitor cocktail. The protein amounts in the lysates were quantified using a BCA assay kit ((Thermo Scientific; 23227). Tissue lysates (2–10 μg) were denatured at 65 °C and resolved by SDS-PAGE (Bio-Rad; #4561033) and then transferred onto polyvinylidene fluoride membranes (Bio-Rad; #1704157) according to the manufacturer's instructions. Membranes were incubated with the primary antibody followed by incubation with the horseradish peroxidase (HRP)-conjugated secondary antibody. The results were visualized using a ChemiDoc Imaging System with a western chemiluminescent HRP substrate (Merck Millipore; WBKLS0100).

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