Figure 2: PHF20 does not affect the upstream signalling events of NF-κB activation.
From: PHF20 regulates NF-κB signalling by disrupting recruitment of PP2A to p65
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.
