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. 2012 Dec 28;24(12):5177–5192. doi: 10.1105/tpc.112.107235

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© 2012 American Society of Plant Biologists. All rights reserved.

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Figure 7. — VICTR Associates in Planta with EDS1, PAD4, and eds1^L262P.

(A) Myc-tagged EDS1, PAD4, and GUS were transiently coexpressed with HA-VICTR in N. benthamiana leaves. Purified nuclear extracts were immunoprecipitated (IP) and immunoblotted (IB) with the indicated antibodies. Top panel shows the pull-down of PAD4 and EDS1, but not GUS, with VICTR. Bottom two panels show input fractions (n = 3).

(B) eds1^L262P does not strongly interact with PAD4 in BiFC assays. Reciprocal BiFC assays of PAD4 with wild-type EDS1 and eds1^L262P were performed by Agrobacterium-mediated transient expression in N. benthamiana. Reconstituted YFP is indicated by yellow fluorescence. Bar values are indicated.

(C) VICTR interacts with eds1^L262P. Co-IP was performed as described above. Top panel is the co-IP of PAD4, EDS1, and eds1^L262P, but not GUS, with VICTR. Bottom two panels show the input fractions. HA-VICTR was detected only after immunoprecipitation with anti-HA antibodies. Migration and sizes (in kilodaltons) of molecular mass standards are at the left of the respective immunoblot panels. Each experiment was performed twice with similar results.