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Figure 1.
Figure 1: Schematic Yap1 structures and in vivo analysis of
Yap1-RD^GFP subcellular localization and oxidation. a, Yap1
contains three conserved regions: a basic leucine zipper DNA
binding domain (bZIP), an n-CRD (Asn279 to Arg313) and a c-CRD,
(Asn565 to Asn650). The NLS and NES are located at the N and C
termini, respectively. The Cys303 -Cys598 and Cys310 -Cys629
disulphide bonds are shown with red lines. The oxidized Yap1-RD
construct used for structure determination consisted of the
protease-resistant n-CRD and c-CRD domains. Yap1-RD^GFP
consisted of an SV40 NLS, GFP and residues Asn279 to Arg313
fused to residues Asn549 to Asn650 of Yap1. This fragment
encompasses the n-CRD and c-CRD sequences plus a small amount of
the native linker. b, Fluorescence microscopy of wild-type and
 gpx3
cells expressing Yap1-RD^GFP from the native YAP1 promoter on a
CEN plasmid. c, Oxidized and reduced Yap1-RD^GFP extracted from
wild-type and gpx3
cells. Exponentially growing cells were either exposed to
H[2]O[2] or left untreated. Cell extracts were run on
non-reducing and reducing SDS -PAGE gels and probed with a GFP
antibody.
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