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. Author manuscript; available in PMC: 2014 Mar 5.
Published in final edited form as: Nat Commun. 2013;4:2062. doi: 10.1038/ncomms3062

Figure 2. PHF20 does not affect the upstream signalling events of NF-κB activation.

Figure 2

HeLa cells were infected with either rAd–Lac or rAd–flag–PHF20 for 24 h, and then cells were treated with TNF for indicated times. (a,b) Whole-cell lysates were immunoblotted with indicated antibodies. (c) Cell extracts were subjected to immunoprecipitation with anti-IKK-γ antibody, and its activity was assessed by immune complex kinase assay as described in Methods (upper panel). The efficiency of immunoprecipitant of IKK complex was confirmed by immunoblotting with anti-IKKγ antibody (lower panel). Relative densities of phosphorylated GST–IκBα were obtained by densitometry for corresponding IKK-γ at each lane. (d) Cell extracts from each sample were subjected to immunoprecipitation with anti-IKKγ antibody. Immunoprecipitates were analysed by immunoblotting with indicated antibodies. A total of 1% of the cell extract volume (WCL) from each sample was used as input control.