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. 2017 Nov 9;292(50):20354–20361. doi: 10.1074/jbc.AC117.000548

Figure 1.

Figure 1.

Development of a KTR for Fus3 activity. A, top panel, diagram of the MAPK signaling pathway that mediates mating differentiation in yeast. Bottom panel, representative images of cells undergoing elongated growth and shmoo formation. B, schematic of the KTR for Fus3 developed in this study. Green regions, NLS; blue regions, NES. C, representative time-lapse images showing the reporter translocation in response to pheromone stimulation. D, average time traces of reporter translocation response to a 30-min pulse of 1 μm pheromone treatment. The reporter response was quantified as the cytoplasmic over nuclear fluorescence intensities (C/N ratio) and was normalized by the basal level. Means and S.E. are shown for WT (blue curve), kss1Δ (red curve), and fus3Δ (green curve). E, average time trace of reporter response in the FUS3as, KSS1 strain (blue curve). Cells were exposed to a constant 1 μm pheromone treatment, and 10 μm 1-NM-PP1 was added 30 min after pheromone stimulation. The response from cells without pheromone exposure was shown as a control (gray curve).