FIG 9.

C. neoformans CDA1 is transcriptionally upregulated during host infection. Mice were inoculated with 10⁷ CFU of KN99 cells. At day 7 PI, the lungs were excised and homogenized, and fungal cells were harvested and used for the isolation of total RNA. Total RNA (0.5 µg) was used for the synthesis of cDNA, which was subsequently subjected to quantitative real-time PCR using CDA1-specific primers. C. neoformans 18S rRNA transcript levels were used as a reference gene. Transcript levels of the respective genes in the cells used as inoculum served as control. Data are the averages for two independent experiments each with three animals per group. Fold expression was calculated for each gene comparing the extent of its upregulation in the lung samples to YPD-grown samples. Significant differences in the expression levels between genes were compared by ordinary one-way ANOVA, followed by Dunnett’s multiple-comparison test. (****, P < 0.0001 comparing fold expression of CDA1, CDA2, and CDA3 in the lung samples to their respective levels in the inoculum sample grown in YPD).