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. 2013 Jul 16;8(7):e69491. doi: 10.1371/journal.pone.0069491

Figure 8. Kinetics of sexual agglutination and polar cell growth in liquid culture.

Figure 8

(A) Mating kinetics of a homothallic wild-type strain (L968) cultured in SSL−N liquid medium. At hourly intervals, the intensity of agglutination (AI) was determined. Portions of the culture were subjected to brief sonication and microphotographs were taken. The long (L) and short (S) axes of cells were measured to calculate the L/S ratio for triplicate samples (100 cells each). The frequencies of prezygotes and cells with pointy projections were determined. (B) Microphotographs showing typical cell morphology. Arrows, prezygotes with a pointy mating projection (shown by asterisks). Scale bar, 10 µm. (C, D) Mating kinetics of M-factor-less mutant cells incubated with (C) or without (D) 200 nM M-factor. The experimental procedures are as described in (A). (E) Left, polar cell growth and prezygote formation in M-factor-induced aggregated cells and floating free cells. An M-factor-less strain (FS55) and an M-factor-producing fus1Δ strain (FS123) were incubated with 200 nM M-factor. After 4 hr, aggregates and free cells were isolated. After brief sonication, the L/S ratio was determined (n=1,000 for both strains). Right, frequency of prezygotes formed with fus1Δ (Aggregates vs Free: each p-value was shown, t-test)..