Fig. 1.

Analysis of interactions of selected members of the SnRK2 family with clade A PP2C phosphatases. a Yeast two-hybrid analysis; growth of yeast expressing the indicated constructs was monitored on selective media: without Leu and Trp (−LW); without Leu, Trp and His (−LWH) supplemented with different concentrations of aminotriazole (3-AT); without Leu, Trp and Ade (−LWA). AD, Gal4 activation domain; BD, Gal4 binding domain. Data represent one of three independent experiments showing similar results. b Pull down assays; GST-fused ABI1 or GST alone, bound to glutathione-sepharose beads, were incubated with recombinant SnRK2.4. Proteins bound to the resin were separated by SDS-PAGE, electroblotted to a membrane and visualized by immunoblotting using anti-SnRK2.4/SnRK2.10 antibodies. In parallel, visualization of proteins in samples used for the Western blotting was performed by staining with Coomassie Brilliant Blue, CBB. Data represent one of two independent experiments showing similar results