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. 2008 Jan 23;105(4):1279–1284. doi: 10.1073/pnas.0706552105

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© 2008 by The National Academy of Sciences of the USA

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Fig. 4. — Differential ubiquitylation of IKKγ alleles. (A) The 293 cells were transfected with plasmids encoding AU1-tagged IKKγ, myc-tagged Ub, and HA-tagged TRAFs. Lysates were precipitated with an antibody against AU1. Precipitates and lysates were blotted for AU1-IKKγ, HA-TRAFs, and myc-Ub. (B) The 293 cells were transfected with plasmids encoding AU1-tagged IKKγ alleles (wild type, K270A, D304N, E308V, and R331P) and HA-tagged TRAF6 (wild type or C70A). Lysates were immunoprecipitated with an antibody against AU1. Precipitates and lysates were blotted for AU1-IKKγ and HA-TRAFs. (C) The 293 cells were transfected with plasmids encoding AU1-tagged IKKγ alleles containing either one mutation (K270A) or two mutations (K270A/D304N, K270A/E308V, and K270A/R331P) and HA-tagged TRAF6 (wild type or C70A). Lysates were immunoprecipitated with an antibody against AU1. Precipitates and lysates were blotted for AU1-IKKγ and HA-TRAFs. The asterisk indicates lanes that were removed electronically from the blot.