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. Author manuscript; available in PMC: 2013 Jul 1.
Published in final edited form as: Cell Microbiol. 2012 Mar 27;14(7):1071–1084. doi: 10.1111/j.1462-5822.2012.01779.x

Fig. 5. Insertion of alanine residues at the PBS1 cleavage site activates RPS5.

Fig. 5

A. Schematic representation of synthetic PBS1 constructs. The kinase domain (amino acids 78-360) and activation segment (amino acids 231-260) of PBS1 are represented by a light blue box and dark blue box, respectively. Synthetic PBS1 constructs containing the indicated alanine insertions after lysine 243 are shown above the diagram.

B. HR phenotypes of Agrobacterium-infiltrated N. benthamiana leaves. The indicated synthetic and wild type PBS1:HA constructs were transiently co-expressed with RPS5-Myc or with RPS5-Myc and AvrPphB. 50 μM dexamethasone was sprayed 40 hours after injection and photographs were taken 24 hr later. This experiment was repeated three times with similar results.

C. Quantification of the HR triggered by the PBS1 constructs described in (B) using electrolyte leakage. Error bars indicate standard deviation. Conductivity was measured at the indicated time points after dexamethasone treatment. This experiment was repeated once with a similar result.

D. Expression of PBS1 constructs described in (B) was confirmed by immunoblot analysis with anti-HA antibody. The same blot was stained with Ponceau S solution to show equal loading of protein samples. Samples were taken 4 hr after dexamethasone treatment.