Nitric Oxide Production in RAW264.7 Cells

RAW264.7 cells were seeded and grown at 37°C and 5% CO₂ for 24 h. RAW264.7 cells were cultured in 1 μg/mL dexamethasone (Sigma, United States), 10–300 μg/mL PLE and 500 ng/mL lipopolysaccharide (LPS) for 24 h. Then, cell supernatants were mixed with Griess reagent (1:1 ratio), and the NO concentration was measured at 540 nm (Lee et al., 2019 (link); Jo et al., 2020 (link)). Cell viability was analyzed using Ez-Cytox reagent (DoGenBio, Korea) according to the manufacturer’s protocols (Yu et al., 2020 (link); Jo et al., 2023a (link)). This experiment was repeated triplicated.

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Jo H.G., Baek C.Y., Kim D., Kim S., Han Y., Park C., Song H.S, & Lee D. (2024). Network analysis, in vivo, and in vitro experiments identified the mechanisms by which Piper longum L. [Piperaceae] alleviates cartilage destruction, joint inflammation, and arthritic pain. Frontiers in Pharmacology, 14, 1282943.

Publication 2023

dexamethasone Ez-Cytox reagent

Corresponding Organization : Gachon University

Other organizations : Kyung Hee University, National Development Institute of Korean Medicine

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Variable analysis

independent variables

Dexamethasone (1 μg/mL)
PLE (10–300 μg/mL)
Lipopolysaccharide (LPS, 500 ng/mL)

dependent variables

NO concentration (measured at 540 nm)
Cell viability (analyzed using Ez-Cytox reagent)

control variables

RAW264.7 cells were seeded and grown at 37°C and 5% CO2 for 24 h

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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