TNF-α, IL-1β, and IL-6 produced by LPS-activated RAW 264.7 cells were determined by using mouse TNF-α, IL-1β, and IL-6 ELISA kit, respectively. RAW 264.7 cells were incubated at 5 × 103 cells/well in 96 well-plates for 24 h at 37 °C and 5% CO2. And then the cells were treated with various concentrations (0.3125, 0.625, 1.25, 2.5, 5, and 10 μM) of 3-CT in the presence of LPS (1 μg/mL). The cells without treatment served as the blank control. The cells with LPS + DMSO (final concentration of 0.1% (v/v)) in the absence of 3-CT served as the negative control (LPS group). After 24 h treatment, TNF-α, IL-1β, and IL-6 were analyzed following the manufacturer’s protocols.
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