ATP was measured using a bioluminescence assay kit (Beyotime Biotechnology, Shanghai, China). Briefly, the fresh kidney samples were lysed in the lysis buffer provided with the kit. The supernatant was collected by centrifugation at 12,000 rpm for 5 min at 4 °C. The concentration of ATP present in the samples was determined by mixing 20 μL of the supernatant with 100 μL of luciferase reagent; the luciferase present in the reagent catalyzes the production of luminescence from ATP and luciferin. The luminescence of each sample was measured on a microplate luminometer (Synergy Mx, BioTek Instruments Inc., Winooski, VT, USA). A standard curve of ATP was prepared using a series of standards of known concentrations; the measured ATP is presented as nmol/mg of protein.

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