Figure 9.

The HD2A and HD2C physically interact with each other. (A) Y2H assays showing interactions of HD2A and HD2C at the protein level. Transformed yeast cells were plated on minimal selective medium DDO (double dropout) and sub-cultured to QDO (quadruple dropout) and QDO/X/A (QDO supplemented with X-alpha galactosidase and Aureobasidin-A) medium to determine their ability to grow to identify protein interactions. Empty vector pGBKT7 was co-transformed with either HD2A-AD or HD2C-AD as a negative control. Blue colonies indicate protein-protein interaction. Three independent experiments were performed in this test. (B) BiFC (Bimolecular fluorescence complementation) assays showing interactions of HD2A and HD2C in the plant host. The constructs HD2A-YN and HD2C-YC (or HD2A-YC and HD2C-YN) were co-infiltrated into leaf epidermal cells of Nicotiana benthamiana via for transient expression. YFP (Yellow Fluorescent Protein) signal was detected after 48-72 hours of infiltration. Empty vector pEarleyGate202-YC was co-transformed with either HD2A-YN or HD2C-YN as a negative control. Three independent experiments were performed in this test. (scale bar 50 µm).