Chemicals and kits
Cell culture reagents were purchased from Thermo Fisher Scientific (Waltham, MA, U.S.A.). Protein assay kit was purchased from Bio-Rad (Hercules, CA, U.S.A.). TNF-α, IL-10, and IL-1β immunoassay kits were purchased from eBioscience (San Diego, CA, U.S.A.). The kits were used according to manufacturer´s instructions. Routine reagents and LPS fromEscherichia coli (O111:B4, L2630) were purchased from Sigma Chemicals (St. Louis, MO, U.S.A.) and recombinant α-Klotho from R&D Systems (1819-KL-050, Minnesota, MN, U.S.A.). All solutions were prepared immediately before use.