FIG. 4.
Expression of eng2+ is essential for the release of ascospores from asci. The haploid eng2Δ strain carrying Pnmt1-eng2 (OL958) and the haploid strains from the opposite mating type, OL176 (WT) and OL773 (eng2Δ), were grown on YES medium to mid-log phase. Equal numbers of cells were collected and spotted onto EMM-N plates with (+T) and without (−T) thiamine to induce mating and sporulation. At the indicated times, aliquots were collected for microscopic inspection and quantification of the percentage of free spores. The crosses were eng2+/eng2Δ (WT), eng2Δ/eng2Δ (eng2Δ), and eng2Δ/eng2Δ plus Pnmt1-eng2 (HA-eng2). (A) Western analysis of Eng2-HA after 24 of incubation in EMM-N medium. (B) Percentages of free spores. (C) Sample images of the different strains grown in the presence and absence of thiamine.