Figure 8.

Tpk1 and Tpk2 are involved in the regulation of virulence-related gene expression. Cells were grown overnight in YPD at 30°C, washed twice with dH₂O, diluted to 0.1 OD₆₀₀ in 10 mL of Spider medium, and incubated for 4 h at 37°C. Total RNA was extracted using TRIzol and treated with the TURBO DNA-free™ Kit to remove genomic DNA. cDNA synthesis was carried out using 200 ng of RNA with the high-capacity cDNA reverse transcription kit. Transcript expression levels were analyzed by SYBR® Green PCR Master Mix and normalized to the C. tropicalis ACT1 gene using $2^{-\Delta \Delta \text{Ct}}$ method. Asterisks indicate statistically significant differences compared with the wild type using Dunnett's multiple comparison test compared to wild type (*P < 0.05; **P < 0.01; ***P < 0.001).