FIGURE 1.

WTX directly interacts with KEAP1. A, GLUE-WTX interacts with endogenous KEAP1. HEK293T parental or HEK293T cells stably expressing GLUE-WTX were treated with DMSO, 100 μm tBHQ, or 100 μm tBHQ and WNT3A conditioned media (WNT3ACM) for 2 h. Lysates were subjected to streptavidin affinity pulldown (AP) followed by Western blot (WB) analysis. B, WTX directly binds KEAP1. Recombinant GST-VSV-WTX was incubated with recombinant CUL3 or KEAP1. After GST affinity purification, complexes were washed, and associated proteins were resolved by Western blot. * denotes full-length GST-VSV-WTX. C, WTX interacts with the KELCH repeats of KEAP1. HEK293T cells stably expressing GLUE-WTX were transfected with FLAG-tagged KEAP1 constructs. Lysates were subjected to streptavidin affinity pulldown followed by Western blot analysis. IB, immunoblot.