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Determination the Production of Nitric Oxide
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To analyze the production of nitric oxide, RAW264.7 cells in 6-well plates were cultured for 3 h with or without C + R + I and then incubated with LPS for 24 h. NO production was then evaluated by quantifying the nitrite in the culture medium using the Griess reagent. Briefly, 100 μL of the collected supernatant was mixed with Griess reagent and incubated at room temperature for 15 min. Absorbance values at 540 nm were recorded, and the levels of nitric oxide were determined according to the standard curve of sodium nitrite ranging from 0.1 to 10 μM.
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