(A) C. neoformans
a-α bilateral crosses of the wild-type, ssn8-1 mutants and SSN8 overexpressed strains were conducted on V8 agar medium in the dark and cells were collected at indicated time for RNA preparation. The abundance of the MFα (a) and GPA2 (b) transcripts were measured by quantitative real time PCR analysis. (B) C. neoformans strains were grown on FA medium and cells were harvested at 0 (a) and 48 (b) h for RNA extraction. The expression of MFα was detected by northern blot analysis and RNA loading for each sample was shown by rRNA staining. Samples in lane 1 and 2 (MATα wild-type), lane 3 and 4 (MATα ssn8-1), lane 5 and 6 (MATα ssn8-2), lane 7 and 8 (SSN8 overexpression strain), lane 9 and 10 (SSN8 reconstituted strain) are as indicated. (C) C. neoformans MATα wild-type, ssn8 mutants, reconstituted strain, and SSN8 overexpression strain were grown in YPD liquid medium for 22 h. Cells were harvested and subjected to RNA extraction. Expression of the GPB1, GPA2, MFα and Sxi1α genes was measured by quantitative real time PCR analysis.