Skip to main content
. Author manuscript; available in PMC: 2016 Jan 31.
Published in final edited form as: Exp Cell Res. 2014 Oct 7;331(1):58–72. doi: 10.1016/j.yexcr.2014.09.034

Figure 3. NF-κB inhibition by Withaferin A is upstream of IKK phosphorylation, but downstream of TAK1 activation.

Figure 3

(A) HEK293 cells were treated with TNF at 10 ng/ml and pre-treated with WA at the dose indicated 30 minutes prior to TNF treatment. Cells were lysed and assayed by western blot.

(B) HEK293 cells were treated with 5 µM WA or 10 ng/ml TNF at the time course indicated. Cells were lysed and immunoblotted as indicated.

(C) IKK in vitro kinase assay. HEK293 cells were pre-treated with WA for 30 minutes and treated as indicated. Cells were lysed and the IKK complex was immunoprecipitated using an α-NEMO antibody. Kinase activity was measured using GST-IκBα substrate and quantified by phosphorimager.