Expression of simA. Expression of simA. (A) Reverse transcriptase PCR (RT-PCR) of simA and a benA loading control performed on RNA isolated from wild-type vegetative hyphae grown for 2 days in liquid medium at 25°C, asexual development (conidiation) cultures grown for 7 days on solid medium at 25°C, and yeast cells grown for 6 days in liquid medium at 37°C. A simA transcript could not be detected during vegetative hyphal growth at 25°C. simA is expressed during asexual development at 25°C and highest during yeast growth at 37°C. Relative intensity values, adjusted using the loading controls, are indicated below the lanes. (B) RNA was isolated from wild-type T. marneffei incubated in macrophage medium alone for 24 h at 37°C (-Φ) or from infected LPS-activated J774 murine macrophages 24 h postinfection at 37°C (+Φ). The amount of simA transcript was increased in cells isolated from infected macrophages. Relative intensity values, adjusted using the loading controls, are indicated below the lanes. Kylie J. Boyce et al. mSphere 2018; doi:10.1128/mSphere.00056-18