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. 2006 Aug 3;25(15):3605–3617. doi: 10.1038/sj.emboj.7601243

Figure 7.

Figure 7

(A) COS1 cells were transfected with expression vectors for Keap1-CBD and HA-Nrf2 and the Keap1:Nrf2 complex isolated on chitin beads. The beads were incubated with the indicated peptides, washed, and proteins that remained bound to the beads were analyzed by immunoblot with anti-HA and anti-CBD antibodies. The amounts of peptides added to each sample were 10 ng (lanes 3, 7, 11), 100 ng (lanes 4, 8, 12), 1 μg (lanes 5, 9, 13), and 10 μg (lanes 6, 10, 14). (B) HEK 293 T cells were transfected with expression vectors for the indicated Keap1 and Nrf2 proteins. Total cell lysates were analyzed by immunoblotting with anti-HA and anti-CBD antibodies (bottom two panels). Equivalent amounts of cell lysates were incubated with chitin beads, washed, and proteins that remained associated with the chitin beads were analyzed by immunoblotting with anti-HA and anti-CBD antibodies (top two panels). (C) Reporter assays were performed as described in Figure 5B.