Fig 1.

RT-PCR analysis of VMA3 expression. RNA was extracted from THE1-CIp10 and tetR-VMA3 cells after 6, 18, and 24 h of incubation in unbuffered YPD with doxycycline (DOX). For comparison, RNA was also extracted from THE1-CIp10 and tetR-VMA3 cells incubated for 6, 18, and 24 h in unbuffered YPD without doxycycline. Reverse transcriptase PCR (RT-PCR) was used to amplify the 300-bp VMA3 transcript using the primers RT-VMA3-5Det and RT-VMA3-3Det. After 6 and 18 h of treatment with doxycycline, the VMA3 transcript was still present in the tetR-VMA3 strain. After 24 h of treatment with doxycycline, the VMA3 gene was completely repressed.