Skip to main content
. 2018 Jul 30;41(8):762–770. doi: 10.14348/molcells.2018.0005

Fig. 2. gAd-induced IκBα degradation is mediated by IKK and Akt activation.

Fig. 2

(A) RAW 264.7 cells were treated with gAd (5 μg/ml) and LPS (1 μg/ml) for the indicated times. The IKK complex was immunoprecipitated using anti-IKKα antibodies. The kinase activity of IKK was assayed as described in the materials and methods section. (B) RAW 264.7 cells were preincubated with SC-514 (an IKKβ specific inhibitor, 100 μM) and then, stimulated with gAd (5 μg/ml) for 30 min and LPS (1 μg/ml) for 10 or 30 min in the presence or absence of SC-514. (C) Cells were transiently transfected with DN-IKKβ (K44A) and control plasmid vector (C.V.). At 48 h after transfection, cells were incubated with gAd (5 μg/ml) for 30 min. (D) Cells were treated with gAd (5 μg/ml) for the indicated times. (E) RAW 264.7 cells were pretreated with LY294002 (40 μM) for 2 h and then stimulated with gAd (5 μg/ml) for 30 min. (F) Cells were transfected with DN-Akt and control plasmid vector. At 48 h after transfection, cells were incubated with gAd (5 μg/ml) for 15 or 30 min. Total cellular extracts were subjected to Western blot analysis for p-IκBα, IKKα, IκBα, p-Akt, Akt, and actin. Results are representative of three separate experiments. K.A., kinase activity assay.