FIGURE 3:

PIF-Citrine recruitment to PhyB is fast and reversible. (A) Recruitment of PIF-tagged protein (channel, PIF-mCitrine) to multiple PhyB anchors goes to completion in seconds. Top, PIF recruitment to PhyB-CAAX (plasma membrane); bottom, PIF recruitment to PhyB-HTB2 (nucleus). Cells were exposed to 750-nm light for 3 min before switching to 650-nm light. (B) Dissociation of PIF-mCitrine from PhyB anchors also goes to completion in seconds (channel, PIF-mCitrine). Top, PIF release from PhyB-CAAX (plasma membrane); bottom, PIF release from PhyB-HTB2 (nucleus). Cells were exposed to 650-nm light for 3 min before switching to 750-nm light. (C) Representative recruitment profile of PIF to the PhyB-HTB2 anchor. (D) Fluorescence images of PIF recruitment to PhyB-CAAX with the 30-s time interval (channel, PIF-mCitrine). (E, F) Single-cell recruitment (top) and dissociation (bottom) profiles of PIF to/from the PhyB-CAAX anchor. Shown are the fluorescence intensity on plasma membrane (E) and the ratio between the fluorescence intensity on plasma membrane and in cytosol (F) during PIF association to and dissociation from the PhyB-CAAX anchor, respectively.