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. Author manuscript; available in PMC: 2014 Nov 7.
Published in final edited form as: Plant J. 2013 Dec 9;77(2):235–245. doi: 10.1111/tpj.12381

Figure 3. HopF2, AvrPto and AvrPtoB interact with BAK1.

Figure 3

(A) HopF2 and AvrPto/B interact with BAK1 in Arabidopsis protoplasts. The α-HA co-IP was performed with protoplasts co-expressing FLAG-tagged BAK1 and HA-tagged AvrPto, AvrPtoB or HopF2, and the immunoprecipitated proteins were analyzed in Western blot with an α-FLAG antibody. HopF2 (B) and AvrPto (C) interact with BAK1 in Arabidopsis plants. pBAK1::BAK1-GFP transgenic seedlings with DEX-inducible effector transgene were treated with 5 μM DEX for 12 hr and subjected to an α-GFP co-IP assay, and the immunoprecipitated proteins were analyzed in Western blot with an α-HA antibody. (D) The BiFC assays for HopF2-BAK1 or AvrPto-BAK1 interactions in Arabidopsis protoplasts. The various BiFC constructs were transfected into protoplasts and the fluorescence were visualized under a confocal microscope. Bar=50 μm. (E) The pull-down assays for HopF2-BAK1 or AvrPto-BAK1 interactions. GST, GST-AvrPto and GST-HopF2 were expressed individually in E. coli, purified with glutathione agarose, and used to pull-down the proteins from protoplasts expressing FLAG-tagged BAK1. The pull-downed proteins were analyzed in Western blot with an α-FLAG antibody.