Figure 8.

PD1 interacts with PEP-β. (A) Yeast two-hybrid assays. The predicted mature PD1 protein fused to the GAL4 DNA-binding domain (BD) was expressed in combination with PEP-β proteins fused to the GAL4 activation domain (AD) in yeast strain AH109. Cells were grown on different selective media. Empty BD and AD vectors served as negative controls. -LT, SD medium without Leu and Trp; -HLTA, SD medium lacking His, Leu, Trp, and Ade. X-α-Gal was added to -HLTA plate to indicate the expression of the MEL1 reporter gene. +, Positive control expressing pGADT7-T and pGBKT7-53; −, Negative control expressing pGADT7-T and pGBKT7-Lam. (B) Coimmunoprecipitation assay. Total leaf protein extracted from two-week-old wild type and transgenic seedlings expressing PD1-HA tag fusion protein received CoIP assay by employing Anti-HA-tag mAb-Magnetic Beads. The samples of input, flowout, and elute were examined by specific antibodies for HA and PEP-β, respectively. The asterisk shows a nonspecific band.