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. 2014 Aug 4;33(19):2261–2276. doi: 10.15252/embj.201489468

Figure 4. CRR-B and, to a lesser degree, CRR-A are crucial for iron detoxification but not adaptation to iron starvation.

Figure 4

  1. Schematic view of the HapX Cys and domain organization including comparisons of HapX orthologs from A. fumigatus, A. nidulans, C. albicans, S. pombe, Ustilago maydis Yap1, and S. cerevisiae Yap5.
  2. Strains were grown for 48 h at 37°C on agar plates with the given iron concentration.
  3. Production of biomass during iron starvation (−Fe), iron sufficiency (0.03 mM, +Fe), and iron excess (3 mM, hFe), as well as production of siderophores under iron starvation was monitored after liquid growth for 24 h at 37°C. The data represent the mean ± SD of biological triplicates; the values were normalized to that of strain hapXR carrying a non-mutated S-tagged hapX. Statistically significant differences compared to hapXR are shown in red (two-tailed, unpaired t-test; P < 0.05). Original data with standard deviations are given in Supplementary Table S1.
  4. Northern blot analyses were performed after liquid growth for 24 h at 37°C under iron limitation (−Fe) or after a subsequent 1-h shift into iron sufficiency (sFe). rRNA is shown as a control for RNA quantity and quality.
  5. Western blot analyses were performed after liquid growth for 24 h at 37°C under iron limitation using antisera recognizing the S-tag for detection of HapX, or porin as control for loading. We were unable to detect S-tagged HapX during iron sufficiency or high-iron conditions with this method (data not shown).

Data information: For simplicity, only one mutant per CRR is shown, the respective, phenotypically identical second mutant is shown in Supplementary Fig S3. Strains are derivatives of A. fumigatus AfS77.