4.4. Enzyme-Linked Immunosorbent Assay (ELISA)

The cells (6 × 10⁴ cells/well in 6 well plates) were pretreated with lycopene (0.1, 0.2 μM) or TAK242 (0.5 μM) and stimulated with HDM (20 μg/mL) for 16 h. The culture supernatants were centrifuged at 10,000× g for 5 min and then collected to measure IL-6 and IL-8 levels. The latter were determined using an enzyme-linked immunosorbent assay (ELISA) kit (R&D Systems, Inc., Minneapolis, MN, USA) following the manufacturer’s instructions.