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In vitro IKK assay was performed as previously described [4]. IKK complex was immunoprecipitated using IKKβ antibody from stimulated HAoECs for 3 h at 4 °C. IKKβ immunoprecipitation beads were incubated with 200 μmol/L adenosine triphosphate (ATP) and 1 μg recombinant GST-IκBα (1–55) protein. IKK kinase activity was determined via immunoblot of phospho-serine32/36 of IκBα.
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The Author(s), under exclusive licence to Springer Science+Business Media, LLC part of Springer Nature
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