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. Author manuscript; available in PMC: 2012 Sep 1.
Published in final edited form as: Nat Immunol. 2011 Aug 7;12(9):844–852. doi: 10.1038/ni.2080

Figure 5. The kinase activity of IKKi is required for IL-17-induced Act1 phosphorylation.

Figure 5

A. Immunoblot analysis of Act1, IKKi and GAPDH in lysates from wild type and IKKi-deficient airway epithelial cells untreated or treated with IL-17A (50 ng/ml) or IL-17F (50ng/ml) for indicated times.

B. Lysates from 293HEK cells transfected with Act1+pcDNA3, Act1+ IKKi WT, Act1+K38A IKKi mutant or pcDNA3 were untreated or treated with phosphatase (CIP, 1 h at 37°C), followed by immunoblot analysis with antibodies against Act1, IKKi and GAPDH.

C. Lysates from wild-type or Act1-deficient MEFs untreated or treated with IL-17 (50 ng/ml, 20 min) were immunoprecipitated with anti-Act1 followed by in vitro kinase assay with or without recombinant IKKi-GST.

D. Lysates from wild-type MEFs or wild-type and Act1-deficient kidney epithelial cells untreated or treated with IL-17 (50 ng/ml, 0, 15 and 30 min) were immunoprecipitated with anti-IKKi followed by in vitro kinase assay.

E. Immunoblot and real-time PCR analysis of IKKi and GAPDH in lysates from wild type and IKKi-deficient airway epithelial cells untreated or treated with IL-17A (50 ng/ml) for indicated times.

The data shown in this figure are representation of three independent experiments.