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. 2005 Dec 15;25(1):255–265. doi: 10.1038/sj.emboj.7600910

Figure 1.

Figure 1

Adi3 autophosphorylation and protein interactions. In (A) and (C), the top panel shows the kinase assay (phosphorimage) and the bottom panel the assay input (Coomassie-stained gel). (A) Adi3 encodes a functional protein kinase. Analysis of kinase-active, -deficient, and constitutively active MBP-Adi3 fusion proteins by in vitro autophosphorylation assays is shown. (B) Adi3 interactions in the Y3H assay. The indicated bait and prey constructs were tested in the Y3H assay (Bogdanove and Martin, 2000) for expression of the lacZ gene on X-Gal plates (blue=interaction). The Drosophila proteins Bicoid and Dorsal were used as negative controls and indicate that the Pto/Adi3/AvrPto interaction is specific. + indicates the presence of AvrPto. (C) Analysis of cross-phosphorylation between Adi3 and Pto. Kinase-deficient MBP-Adi3 proteins were used as substrates for kinase-active MBP-Pto protein with or without AvrPto-FLAG protein. Kinase-deficient MBP-PtoD164N protein was used as a substrate for kinase-active MBP-Adi3 with or without AvrPto-FLAG protein. Molecular weight (kDa) markers are indicated.