(A) Mating phenotypes for wild type XL280α, two independent prm1 mutants (CF317 and CF659), and two independent kar5 mutants (CF150 and CF260) during C. deneoformans unisexual reproduction. The scale bars are 100 μm and 20 μm for top row and bottom row, respectively. (B) Gene expression patterns for PRM1, KAR5, and MFα were examined by RT-PCR (* indicates p <0.05 and ** indicates p <0.005 for each pairwise comparison). Wild type (XL280α) was grown on YPD medium for 36 hours, and on V8 medium for 36 hours or one week. prm1 mutant (CF317) and kar5 mutant (CF150) were grown on V8 medium for 36 hours. The error bars represent the standard deviation of the mean for the three biological replicates. (C) Comparison of wild type cell-cell fusion frequency among three different sexual cycles between C. neoformans and C. deneoformans. (D) Unilateral and bilateral prm1 mutant and kar5 mutant cell fusion frequency compared to wild type. (E) Scanning electron microscopy of basidium morphology and sporulation patterns (indicated by arrows) of the wild type (XL280α) and the kar5 mutant (CF150). The scale bar is 5 μm.