Figure 2 - MUC1-CD binds directly to IKK and IKK.

(a) Lysates from the indicated cells were subjected to immunoprecipitation with a control IgG or anti-IKK antibody. The precipitates were immunoblotted with the indicated antibodies. (b) GST and GST–IKK bound to glutathione–agarose beads were incubated with purified MUC1-CD. The precipitates and input were immunoblotted with an anti-MUC1-C antibody. Input of the GST and GST–IKK proteins was assessed by Coomassie blue staining. (c) Amino-acid sequence of MUC1-CD (upper panel). GST and the indicated GST–MUC1-CD fusion proteins bound to glutathione beads were incubated with purified IKK. The precipitates and input were immunoblotted with an anti-IKK antibody (lower left). GST and the indicated GST–IKK fusion proteins bound to glutathione beads were incubated with MUC1-CD. The precipitates and input were immunoblotted with an anti-MUC1-C antibody (lower right). Input of GST and GST– fusion proteins was assessed by Coomassie blue staining. (d) Lysates from the indicated cells were subjected to immunoprecipitation with a control IgG or an anti-IKK antibody. The precipitates were immunoblotted with the indicated antibodies. (e) GST and GST–IKK bound to glutathione beads were incubated with purified MUC1-CD. The precipitates and input were immunoblotted with an anti-MUC1-C antibody. (f) GST and the indicated GST–MUC1-CD fusion proteins bound to glutathione beads were incubated with purified IKK. The precipitates and input were immunoblotted with an anti-IKK antibody. Input of the GST and GST– fusion proteins is shown in Fig. 2c, left. (g) GST and the indicated GST–IKK fusion proteins bound to glutathione beads were incubated with MUC1-CD. The precipitates and input were immunoblotted with an anti-MUC1-C antibody. Full scans of the gels in a, b, c, e and f and g are shown in Supplementary Fig. S6-2.