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.