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. 2010 Jul 9;5(7):e11514. doi: 10.1371/journal.pone.0011514

Figure 3. Sexual differentiation in PKA mutants overexpressing Tor2.

Figure 3

(A and B) Cells of the opposite mating types of wild-type, cyr1Δ, nmt-tor2 and nmt-tor2 cyr1Δ were grown to mid-exponential phase, spotted onto malt extract plates and incubated for 24 hours at 25°C. (A) Percentage of zygotes, (B) Iodine staining. Tor2 overexpression impaired sexual differentiation. This phenotype was rescued in a cyr1Δ background. (C and D) Wild-type, cyr1Δ, nmt-tor2 and nmt-tor2 cyr1Δ cells were grown to mid-exponential phase in minimal medium at 25°C, washed several times and incubated in minimal medium lacking nitrogen. Cells were collected at the indicated times and RNA and protein were extracted. (C) Northern blot hybridized with a probe against ste11+ gene. (D) Western blot using anti-Ste11 antibody. Cells overexpressing Tor2 showed lower ste11+ mRNA and protein levels than the wild-type. ste11 mRNA and protein levels are clearly increased in cells lacking Cyr1 (nmt-tor2 cyr1Δ). rRNA and tubulin were used as loading controls.