FIG 1 .

PCK1 expression does not correlate with CNS persistence of C. neoformans. (A) PCK1 expression was measured by real-time PCR following exposure to either CSF or YNB-0.05% glucose (low glucose) for 24 h at 37°C. PCK1 expression following growth in YPD at 37°C (in vitro 37°C) was used as a positive control. (B) PCK1 expression in C. neoformans was measured by real-time PCR during rabbit infection. Expression levels in vivo were compared to those from yeast grown in YPD at 37°C (in vitro 37°C). (C) PCK1 expression levels were measured by real-time PCR for C. neoformans isolated from human CSF samples. C. neoformans strains 44-1 and HC1 were obtained from the Duke University Infectious Disease Specimen Repository, which houses specimens isolated from deidentified patients with cryptococcal meningitis. PCK1 transcripts for C. neoformans cultured from the CSF samples and grown in YPD at 37°C (“in vitro” samples) were compared to transcripts isolated directly from the yeast in CSF (“in vivo” samples). (D) Levels of persistence of the WT, acs1∆, snf1∆, or snf1∆ SNF1 mutant strains of C. neoformans were compared by measuring the number of CFU of each strain isolated from the CSF of male NZW rabbits over time as described previously (24, 42). Rabbits were infected with 10⁸ CFU of either the WT, the acs1∆, the snf1∆, or the snf1∆ SNF1 strain, and fungal burden was assessed over the course of 2 weeks as described in Materials and Methods. Missing data for the snf1∆ SNF1 strain on day 13 reflects the mortality of all these rabbits before day 13. Statistical differences were assessed by an ANOVA comparison of the graphed lines using the fit model process in JMP version 8 (SAS Institute, Inc., Cary, NC) (*, P < 0.0001).