RAW264.7 cells were pre-treated with Sb-EE (0, 25, 50, 100, 150 and 200 µg/mL) for 30 min, then stimulated with LPS (1 µg/mL) for 24 h. The supernatant (100 μL) was mixed with 100 μL of Griess reagent, as reported previously (Hossen et al. 2017; Kim et al. 2018). Griess reagent contains 0.2% naphthylethylenediamine dihydrochloride (NEDD) and 2% sulphanilamide in 5% phosphoric acid. The absorbance of this mixture was measured at 540 nm and the concentration of nitric oxide (NO) was calculated by comparison with a standard curve.

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