Ua assay kit

Mouse urine was collected for 24 h employing metabolic cages and preserved at −80 °C. Urine albumin, serum IgA, and urine and serum creatinine were quantified using an albumin ELISA Kit (WAKO, Osaka, Japan), mouse IgA ELISA Kit (Crystal Chem, IL USA), and creatinine ELISA Kit (WAKO, Osaka, Japan), respectively. All assays were performed according to the manufacturers’ protocols. Serum UA concentrations were determined with a UA assay kit (Cayman Chemical, MI), according to the manufacturer’s instructions.

Mouse urine was collected for 24 h employing metabolic cages and preserved at −80 °C. Urine albumin and creatinine were quantified using a mouse albumin ELISA kit (FUJIFILM Wako Shibayagi, Gunma, Japan) and a mouse creatinine assay kit (Abcam, Cambridge, MA, USA), respectively. Serum UA concentrations were assayed with a UA assay kit (Cayman Chemical, Ann Arbor, UI, USA). All assays were conducted according to the manufacturers′ protocols. The albumin-to-creatinine ratio (ACR) was calculated as urinary albumin/urinary creatinine.

The uricase inhibitor PO, adenine (AD), and XOD inhibitor allopurinol (AP, urate lowering agent) were purchased from Sigma (St. Louis, MO, United States). TGF-β was obtained from R&D Systems, Inc. (Minneapolis, MN, United States). The NLRP3 inhibitor MCC950, UA assay kit and XOD fluorometric assay kit were purchased from Cayman Chemical Company (Michigan, United States). Pterostilbene (96% purity) was a gift from Sabinsa Corporation (East Winsor, NJ, United States). Primary antibodies against E-Cadherin, fibronectin, α-SMA, and Vimentin were purchased from Genetec Inc. (Montreal, QC, Canada). Antibodies against NLRP3, ASC, caspase-1, IL-1β, AMPK, phosphor-AMPK, mTOR, phosphor-mTOR, p62, LC3-II, and GAPDH were purchased from Cell Signaling (Beverly, MA, United States). The horseradish peroxidase (HRP)-conjugated anti-mouse and anti-rabbit secondary antibodies were obtained from Jackson ImmunoResearch (West Grove, PA, United States).