Fig 2. Expression levels of genes involved in carbon metabolism were analyzed in the acuK and acuM deletion mutants.
The 108 conidia/ml of T. marneffei wild type, ΔacuK, and ΔacuM strains were precultured in the minimal medium containing glucose for 16 hours before transferring into acetate, ethanol and proline gluconeogenic carbon source medium. After incubation at 25 °C for 24 hours, the fungal cells were collected and subjected for RNA extraction and cDNA synthesis. The qRT-PCR was used to determine the fold-change of gene transcripts (A) fbp (fructose-1,6-bis phosphatase), (B) prnD (proline dehydrogenase), (C) facB (key regulator in acetate utilization), and (D) alcB (alcohol dehydrogenase). Gene expression levels were calculated by the 2-ΔΔCt method using actin as a reference gene. Error bars indicate standard deviation. Statistical analysis was performed using unpaired-t-test (GraphPad Prism version 7.00 for Windows). Statistically significant values (ns = non-significant, P>0.05, * P≤ 0.05, ** P≤ 0.01, *** P≤ 0.001, **** P≤ 0.0001) are indicated. Experiments were performed in three replicates.