The Candida albicans-Specific Gene EED1 Encodes a Key Regulator of Hyphal Extension
Figure 4
Expression of the UME6 gene depends on EED1 expression levels.
(A) C. albicans eed1Δ mutant cells harboring the ectopic pTET-EED1 were grown on plastic (RPMI1640 medium, 37°C, 5% CO2) for 12 h either in the absence or presence of 50 µg/ml doxycycline (with or without DOX) in the medium. Scale bar: 20 µm. (B) Expression of the UME6 gene was down- regulated in eed1Δ compared to wild type. Expression was restored after overexpression of pTET-EED1. Expression was detected by qRT PCR using SYBR Green. Gene expression was normalised against EFB1 and ACT1. Total RNA was isolated after 12 h growth on plastic surfaces. Asterisk means p≤0.05. (C) Infection of reconstituted human oral epithelium (RHE, Skinethic) was carried out using the eed1Δ mutant harboring the ectopic pTET-EED1 cassette. Induction of EED1 expression was achieved by the addition of 50 µg/ml doxycycline to the infection. Pictures show PAS stained sections of the RHE tissues. (D) Average LDH release caused by eed1Δ + pTET-EED1 exhibited in the presence and absence of doxycycline in three independent infections compared to uninfected RHE tissue.