Fig. 2: TaSG-D1 and TaSG-D1^E286K interact with TaPIF4.

a TaSG-D1/TaSG-D1^E286K interact with the C-terminus of TaPIF4 in a Y2H assay. 35 mM 3-amino-1,2,4-triazole (3-AT) was used to suppress self-activation in the TaSG-D1^E286K and TaPIF4 interaction assay. SD-L/W stands for media lacking Leucine and Tryptophan. SD-L/W/H/A stands for media lacking Leucine, Tryptophan, Histidine and Adenine. b TaSG-D1/TaSG-D1^E286K interact with TaPIF4 in N. benthamiana leaves in an LCI assay. The experiment was performed five times. c TaSG-D1/TaSG-D1^E286K interact with TaPIF4 in vitro in a pull-down assay. GST/GST-TaPIF4 were incubated with His-TaSG-D1/His-TaSG-D1^E286K. Protein mixture was immunoprecipitated with Proteinlso GST Resin and detected with anti-His (upper panel) and anti-GST (lower panel) antibodies. The arrow in lower panel (~75 KDa) indicates GST-TaPIF4. “**” in the upper panel indicates non-specific band, the arrow in the upper panel indicates non-phosphorylated His-TaSG-D1/His-TaSG-D1^E286K (~48 KDa), “*” in the upper panel indicates self-phosphorylated His-TaSG-D1/His-TaSG-D1^E286K. Three independent experiments were performed. d TaSG-D1/TaSG-D1^E286K interact with TaPIF4 in vivo in a Co-IP assay. 35 S: TaSG-D1-GFP/35 S: TaSG-D1^E286K-GFP/35 S: GFP and 35 S: TaPIF4-Myc were co-expressed in N. benthamiana leaves. Protein extract was immunoprecipitated with anti-GFP magnetic beads and detected using anti-GFP (lower panel) and anti-Myc (upper panel) antibodies. Three independent experiments were performed. Source data are provided as a Source Data file.