Skip to main content
. 2017 Nov 28;12(11):e0188886. doi: 10.1371/journal.pone.0188886

Fig 2. Interaction of PUB12 and PUB13 with CERK1 depends on its kinase activity.

Fig 2

(A) PUB12 and PUB13 interact with the intracellular domain (IC) of CERK1, but not LYK5 in the yeast two hybrid experiment. The growth of yeast colonies on plates (-ULWH) lacking uracil (U), leucine (L), tryptophan (W), and histidine (H) with 2 mM 3-aminotriazole (3-AT) indicates a positive interaction. (B) Immunoblot analysis of yeast extracts containing LexA fusion proteins. Total proteins were prepared from the yeast cells used for the two-hybrid assays in Fig 1B. Proteins were subjected to Phos-tag SDS-PAGE (right) or SDS-PAGE (left). Gels were analyzed by Western blotting with α-LexA. (C) PUB12 and PUB13 interact with CERK1-IC, but not the kinase-inactive mutant (CERK1D441V) in the yeast two hybrid experiment. (D) Co-immunoprecipitation assays show that PUB12-GFP forms a complex with the kinase-active and inactive forms of CERK1 in Nb leaves.