Fig. 1: Yeast two-hybrid assay (Y2H) demonstrates light-dependent interaction between phytochrome and interacting proteins in AH109C.
a Y2H showing red light-dependent interaction between phyBPSM and PIF3. Y2H was performed to observe the red light-dependent interaction between the N-terminal photosensory module of phytochrome B (phyBPSM, amino acids 1-652) and PIF3. Serial dilutions of AH109C yeast cells harboring a GAL4 DNA binding domain (BD)-fused phyBPSM (BD-phyBPSM) with either an empty GAL4 activation domain (AD) vector (AD) or an AD-fused PIF3 (AD-PIF3) were plated on non-selective agar plates lacking leucine and tryptophan (-LW) and selective plates lacking leucine, tryptophan, and histidine (-LWH). The plates were incubated either in the dark (Dc) or under red light (Rc, 15 μmol m−2 s−1). OD600 of 1 was serially diluted (4-folds each). b Red light-dependent interaction between full-length phyB and interacting proteins. The interaction between full-length phyB and its interacting proteins under red light was tested using AH109C cells expressing AD-fused full-length phyB (AD-phyB) and BD-fused interacting proteins (SPA1, SPA2, ELF3, TZP).
