Effect of <i>AFT2</i> deletion on iron content and iron-regulon expression under iron-deficient conditions.

<p>(A) Cells were pre-grown in YPD medium for 24 h at 30°C, and re-cultivated in 100 ml fresh YPD+100 µM BPS medium for 12 h, respectively. Then, cells were washed twice with sterilized PBS buffer, twice with 1 mM EDTA, and followed by twice with distilled deionized water to remove all exogenous iron. Measurement of cellular iron levels was performed by a Hitachi 180-80 polarized Zeeman atomic absorption spectrometry. Note: ppm = mg/kg. (B) Overnight cultures of the wild-type and <i>aft2Δ/Δ</i> mutant strains were cultivated to mid-exponential phase in YPD+100 µM BPS medium, and used for RNA isolation. Quantitative real-time PCR was performed to determine the relative expression changes of iron-responsive genes. Data indicate mean values ± standard deviations from three independent experiments performed in triplicates.