Deletion of IKK2 in hepatocytes does not sensitize these cells to TNF-induced apoptosis but protects from ischemia/reperfusion injury
J. Clin. Invest. Tom Luedde, et al. 115:849 doi:10.1172/JCI23493 [Go to this article.]

Figure 4
A dominant-negative IKK2 form can block NF-κB activation after TNF-α stimulation. (A) IP in 300 μg of whole cell liver protein extracts from Ikk2^f/f and Ikk2Δhepa mice was performed with a polyclonal antibody against NEMO, followed by Western blot analysis with a monoclonal antibody against IKK1 or IKK2 as indicated. Association between NEMO and IKK1 was assessed before as well as 30 and 60 minutes after stimulation with TNF-α. (B) Western blot of protein extracts that were IP with anti-IKK1 and blotted with a monoclonal antibody against IKK1 or IKK2 as indicated. (C) Primary hepatocyte cultures were prepared from livers of Ikk2^f/f and Ikk2Δhepa mice and either treated with PBS alone, infected with LacZAdv, or infected with dnIKK2Adv in a viral dose of 20 PFUs for 12 hours. Protein extracts (200μg) were subjected to IP with a NEMO antibody, followed by Western blot with an IKK2 antibody. (D) Primary hepatocytes from Ikk2^f/f and Ikk2Δhepa mice were infected with LacZAdv or dnIKK2Adv in a viral dose of 20 PFUs for 12 hours and treated with TNF-α (30 ng/ml). Protein extracts were subjected to an NF-κB gel-retardation assay. (E) Protein extracts (100 μg) from primary hepatocytes infected with LacZAdv or dnIKK2Adv were IP with anti-Nemo and blotted with a monoclonal antibody against IKK1.