21-O-Angeloyltheasapogenol E3, a Novel Triterpenoid Saponin from the Seeds of Tea Plants, Inhibits Macrophage-Mediated Inflammatory Responses in a NF-κB-Dependent Manner

Figure 4

Effect of ATS-E3 on the activation of NF-κB upstream signaling cascades. ((a) and (b)) Total and phosphoprotein levels of PDK1, AKT, IKKα/β, and β-actin in whole cell lysates of LPS-treated RAW264.7 cells were determined by immunoblotting analysis. (c) Kinase activities of PDK1 and AKT1 were determined by a direct kinase assay using purified enzymes. The value of the control, which received vehicle treatment, was set as 100% activity for each enzyme (PDK1 or AKT1). (d) NO inhibitory activities of standard compounds (BAY11-7082 (BAY), LY294002 (LY), and wortmannin (Wort)) were determined in RAW264.7 cells (1 × 10⁶ cells/mL) treated with LPS (1 μg/mL) in the presence or absence of standard compounds for 24 h. Relative intensity was calculated using total levels by the DNR Bio-Imaging System. and compared to control group.

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