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Lysates were prepared from TNFα-treated (10 ng/ml, 10 mins) and non-treated transfected HEK 293T/17 cells (American Type-Culture Collection, ATCC CRL-11268) expressing Flag-tagged JNK proteins (Gupta et al., 1996). JNK proteins were isolated by immunoprecipitation using agarose-bound Flag M2 antibody (Sigma-Aldrich Cat# A2220, RRID:AB_10063035) (Gupta et al., 1996). JNK activity was measured using an in vitro protein kinase assay with the substrates cJun and [γ-32P]ATP as substrates (Whitmarsh and Davis, 2001).
Copyright and License information: ©2016, Vernia et al
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