Fig. 5

Characterization of AIP1 protein interactions and subcellular localization. a Schematic representation of AIP1 and ABAP1 proteins. AIP1 harbors two repetitions of Agenet/Tudor (Ag) domains in its N-Terminal and a DUF724 in the C-terminal (DUF); ABAP1 harbors eight Beta-catenin-type Armadillo (ARM) at its N-terminal and one BTB/ POZ (BTB) domain in the C-terminal. b Yeast two hybrid assays with the C-terminal region of AIP1 (aa 540-723) or the complete AIP1 CDS fused with GAL4 DAD (AIP1-C-Term AD and AIP1-CDS AD, respectively) against full-length ABAP1 fused with GAL4 DBD. GAL4 DBD empty vector was used as negative control. Details of the constructs can be found in Additional file 13. Yeast transformation was selected in -L-T (SD medium lacking Leucine and Tryptophan), and protein interactions were selected in -L-T-H or -L-T-H-A (SD medium lacking Leucine, Tryptophan and Histidine, or Histidine and Adenine. c Left: GST pulldown of bacterially expressed recombinant GST-AIP1and HIS-ABAP1. Right: Semi-in vivo pulldown assay of bacterially expressed recombinant GST-AIP1 and protein lysates of Arabidopsis 10-day-old plants. ABAP1 interacting proteins were assayed with antibodies anti-ABAP1 in immunoblots. d Subcellular localization of GFP::ABAP1 and RFP::AIP1 in abaxial epidermis of N. benthamiana 14-day-old leaves by confocal microscopy. RFP::AIP1 inset showing the speckle-pattern in nucleus