Garcinol

Seven-week-old male C57BL/6N mice were obtained from HyoSung Science Inc. (Daegu, Korea) and kept at 20–24 °C and 55% humidity for 1 week. The mice were assigned into 3 groups (n = 8 per group): vehicle (Veh), cisplatin (CP), and cisplatin plus garcinol (CP+Gar). The CP group was given a single intraperitoneal injection of cisplatin (20 mg/kg in 0.9% saline; Sigma-Aldrich, St. Louis, MO, USA). An equal volume of the vehicle was injected intraperitoneally into the Veh group. To investigate the effect of garcinol (Abcam, Cambridge, MA, USA) on cisplatin-induced nephrotoxicity, the CP+Gar group was intraperitoneally injected with garcinol (10 mg/kg) for 4 consecutive days, starting from 1 day prior to cisplatin injection. The doses of garcinol and cisplatin were chosen based on the results of previous studies [29 (link),30 (link),31 (link)]. All mice were sacrificed 72 h after cisplatin injection. All animal experiments were performed in accordance with the Institutional Animal Care and Use Committee of the Daegu Catholic University Medical Center (approval number: DCIAFCR-200626-13-Y).

Recombinant mouse GluOC was prepared as described^{8 (link)}. Myristoylated PKI 14-22 amide (PKA inhibitor) was obtained from Enzo Life Sciences (Farmingdale, NY), U0126 (MEK inhibitor) and AS1842856 (FoxO1 inhibitor) from Merck-Millipore (Darmstadt, Germany), garcinol (p300 inhibitor) and necrostatin-1 (RIP1 inhibitor) from Abcam, staurosporine (inducer of apoptosis) and carvacrol (TRPM7 inhibitor) from Wako (Osaka, Japan), and neutralizing antibodies to FasL from R&D Systems (Minneapolis, MN).