FIGURE 5:

Synergistic activation of Fus3. (A) Dual-phosphorylated Fus3 in wild type and ste5^ND (nondocking) msg5Δ (phosphatase-deficient) mutants replotted from Figure 3 for comparison with the ste5^ND msg5Δ mutant. Dual-phosphorylated Fus3 is quantified as a percentage of total Fus3 at 15 min. Results are reported as ±SEM (n ≥ 3). (B) Transcription reporter data in the same strains as in A, as a percentage of maximum activity in wild type. Inset, Hill slope and EC₅₀ for each strain. Results are reported as ±SEM (n = 3). (C) Pheromone-induced growth arrest for cells treated with α-factor. Halo diameters are quantified for all strains at 5 μg (left). Results are reported as ±SEM (n = 4). Right, representative halo assays for the wild-type and ste5^ND msg5Δ strains at 1.5, 5, and 15 μg. (D) Polarized growth in cells treated with 300 nM α-factor. Representative images for wild-type, msg5Δ, ste5^ND, and ste5^ND msg5Δ cells, each bearing the integrated polar cap marker Bem1-GFP (arrow), at the indicated times. The path of the polar cap was quantified for 10 cells 45–200 min after pheromone addition. Data are representative of two or more experiments. Scale bar, 10 μM.