*3.5. Enzyme-Linked Immunosorbent Assay (ELISA) for PGE2 and TNF-α Measurement*

RAW 264.7 cells (2 × <sup>10</sup><sup>5</sup> cells/well) were plated in 6-well culture plates overnight. In a serum-free medium, cells were pretreated with the methanol extract (1, 10 μg/mL), isolated lignans (5, 10 μg/mL), or indomethacin (10 μM; NSAID) for 3 h and subsequently treated with LPS (5 μg/mL) for 24 h. The supernatants were collected and stored at −80 ◦C until analysis. The assessment of PGE2 and TNF-α levels was quantified with the mouse PGE2 and TNF-α ELISA kits following the manufacturer's instructions.
