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. Author manuscript; available in PMC: 2014 Apr 1.
Published in final edited form as: Cancer Res. 2013 Feb 4;73(7):2199–2210. doi: 10.1158/0008-5472.CAN-12-4400

Figure 2. The ETGE motif is required for KEAP1 association.

Figure 2

(A) Streptavidin affinity purified KEAP1 protein complexes from HEK293T cells were analyzed by Western blot for the indicated FLAG-tagged proteins (SBP, streptavidin binding peptide; HA, hemagglutinin). (B) Streptavidin affinity purified protein complexes from HEK293T cells stably expressing the indicated SBPHA-tagged proteins were analysed by Western blot for the indicated proteins. (C) HEK293T cells stably expressing FLAG-tagged WDR1 or TSC22D4 were transfected with SBPHA-tagged KEAP1, KEAP1 BTB, or KEAP1 KELCH before FLAG affinity purification and Western blot (D) FLAG-tagged wild-type or ET/SGE-deletion constructs were expressed in HEK293T cells stably expressing SBPHA-KEAP1 before streptavidin affinity purification and Western blot. (E) HEK293T cells stably expressing FLAG-tagged KEAP1 were transfected with the indicated expression constructs; associated proteins were revealed by FLAG immunoprecipitation and Western blot. (F) HEK293T cells were transfected with the indicated ET/SGE-containing protein, constitutively expressed Renilla luciferase and the NRF2-driven Firefly luciferase (NQO1-ARE). Error bars represent standard deviation from the mean from 3 biological replicates (* p<0.05; Students T-test).