Measurements of Serum IL-1β, IL-6, and TNF-α

ELISA analyses were performed to quantify the levels of inflammatory cytokines and interleukins in mouse serum. Blood samples were clotted for 2 h at room temperature before centrifugation (4°C, 2000 g, 30 min). The concentrations of IL-1β, IL-6, and TNF-α in the resulting supernatants were measured using a Quantikine ELISA kit (Neobioscience Technology Co, Ltd, Shenzhen, China) following the manufacturer’s protocol. The results are shown as the concentration of cytokine per milliliter of serum. The samples were aliquoted and stored at − 80°C.