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. 2012 Aug 23;8(8):e1002875. doi: 10.1371/journal.ppat.1002875

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© 2012 van Damme et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) An overview of the CRN8 construct, including the catalytic domain (red) and the amino acid positions of Arginine (R) at 469 and Aspartate (D) at 470. (B) The upper panel indicates the protein input for the kinase assay. The lower panel shows an auto-radiogram detecting phosphorylation state of CRN8, CRN8^D470N, and CRN8^R469A;D470A, with only CRN8 producing a signal. (C) Phosphorylated CRN8 protein was detected in crude plant protein extract (top panel) and in FLAG immuno-purified protein (lower panel) by Pro-Q Diamond phosphoprotein in gel stain. The middle panel shows that all three fusion proteins were present, as detected by FLAG antibody on a Western blot. (D) Macroscopic cell death associated with A. tumefaciens-mediated transient expression of FLAG:CRN8 and kinase-inactive mutants FLAG:CRN8^D470N and FLAG:CRN8^R469A;D470A in N. benthamiana. Picture was taken five days post A. tumefaciens infiltration in N. benthamiana.