L1395

Strains used in this work are listed in Table S1. Media were as described in Moreno et al. (1991) (link). Strains were thawed in a solid yeast extract supplemented (YES) rich medium plate and incubated for 24 h at 32°C or 48 h at 25°C and patched in a new fresh plate and incubated for 24 h at 32°C or 25°C. For meiosis induction, strains were repatched in solid sporulation agar (SPA) medium and incubated for 6–6.5 h at 28°C, the optimal temperature for mating and meiosis. After induction, meiotic cells were immobilized with lectin (0.2 µg/ml, Sigma-Aldrich, L1395) at the bottom of a µ-Dish glass-bottom 35 mm uncoated dish (81151, ibidi Gmbh), washed and covered in a total of 3 ml EMM2 minimal medium without nitrogen, with or without drug, to ensure continuation of meiosis.

Channel slides (µ-Slide VI 0.4, ibidi) were treated with lectin (Sigma-Aldrich, L1395) (0.1 mg/mL in water) for ~5 min, washed with YE5S, and cells were added and incubated for ~5 min to allow for attachment. Unattached cells were removed by washing with YE5S. Solutions of BSA (Sigma-Aldrich, A3608) in YE5S were made fresh. Attached cells in the chamber were first imaged in YE5S medium, then shifted transiently to YE5S containing different concentrations of BSA.