Aft2, a Novel Transcription Regulator, Is Required for Iron Metabolism, Oxidative Stress, Surface Adhesion and Hyphal Development in Candida albicans
Figure 7
The enhanced hyphal development of AFT2 deletion is closely associated with increased expression of hypha-specific genes in C.albicans.
(A) Cells were incubated in liquid YPD+10% serum and Spider medium at 37°C, respectively. At the time indicated, the samples were examined and photographed by light microscopy. Hyphal lengths in the wild-type, aft2Δ/Δ mutant, aft2Δ/Δ+Vector (NKF90) and AFT2-overexpression complemented (NKF91) strains were measured, and shown in the graph. (B) After 2 h incubation in liquid inducing medium, cells were harvested and used for RNA isolation. Quantitative expression levels were determined by real-time PCR assays. (C) The strains producing Aft2-HA fusion proteins were incubated for 0 h, 1 h and 2 h in indicated liquid inducing medium, respectively. Then, cells were harvested and used for RNA isolation and protein extraction. Immunoblot analysis was performed using an anti-HA antibody to detect tagged Aft2 proteins. Actin was detected with anti-actin antibody as a loading control.