IKK
β
-Targeted Anti-Inflammatory Activities of a Butanol Fraction of Artificially Cultivated Cordyceps pruinosa Fruit Bodies
Figure 4
Effect of Cp-BF on the upstream signalling for NF-κB activation. (a) Phosphoprotein or total protein levels of IκBα, Akt, p38, ERK, JNK, IKKα/β, and β-actin from cell lysates were determined by phosphospecific or total protein antibodies. (b) Kinase activities of IKKα and IKKβ were determined by a direct kinase assay using purified enzymes. The control was set as 100% for each enzyme activity obtained with vehicle treatment. (c) NF-κB-mediated luciferase activity in IKKβ-transfected HEK293 cells was measured using a luminometer. Phosphoprotein levels of IκBα were determined by immunoblot analysis. (d) Phosphoprotein levels of IκBα, p50, and p65 from IKKβ-transfected RAW264.7 cells were determined by immunoblot analysis. (e) Binding of IKK to IκBα was determined by immunoprecipitation and immunoblot analysis of whole cell lysates of LPS-treated RAW264.7 cells (5 × 106 cells/mL). (f) Level of NO was determined by Griess assay with culture supernatants of RAW264.7 cells treated with BAY11-7082 and LPS (1 μg/mL) for 24 h. The effect of BAY11-7082 on phagocytic uptake of RAW264.7 cells treated with FITC-dextran (1 mg/mL) was determined by flow cytometric analysis. and compared to control.
