Fig. 5.

Western blot analysis and kinase activity assay of IKKα. a RAW 264.7 cells were preincubated for 2 h with indicated concentrations of kinsenoside and then activated for 1 h with RANKL. Nuclear fractions were obtained for the detection of p65 and p50 levels. b RAW 264.7 cells were preincubated for 2 h with indicated concentrations of kinsenoside and then activated for 24 h with RANKL. The whole proteins were obtained for the detection of NFATc1 levels. c Cytoplasmic fractions were obtained for the detection of p-IκBα, IκBα, and p-p65 levels. d Cytoplasmic fractions were obtained for the detection of IKKα, IKKβ, and p-IKKα/β levels. All values are expressed as means ± SD (n = 3). Values not sharing a common superscript differ significantly. e In vitro kinase assay of IKK was done as described in “Materials and methods”. The IC50 values were approximately 57.6 μM. Values were expressed as means ± SD (n = 3)