HMC3 cells were plated into 12-well tissue culture plates (1 × 105 cells/well) and cultured in MEM with 10% FBS for 12 h. Cells were incubated with or without 1000 nM DEX for 30 min, and then stimulated with or without 100 ng/mL LPS for 24 h, based on the previously published protocol [31 (link),32 (link)]. The culture supernatants were then collected and centrifuged at 12,000× g for 10 min, and LDH was measured using the LDH cytotoxicity detection kit according to the manufacturer’s instructions (Takara Bio Inc., Shiga, Japan) using an xMark™ microplate reader (Bio Rad, Hercules, CA, USA).
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