A. fumigatus CrzA transcription factor binds to the promoter regions of specific chitin synthase-encoding genes. (A) Schematic representation of the DNA probes (including mutated probes) of the chsA, csmB, chsC, and chsG promoter regions used during the electrophoretic mobility shift assays (EMSAs). (B) EMSAs using the GST::CrzA-tagged protein. Probes containing the endogenous or mutated CrzA motif were assayed for DNA-binding activity, using the recombinant GST::CrzA protein. The specificity of the DNA-protein binding was confirmed by adding specific competitors (cold DNA probes) and the mutated probes. SC, specific competitor; mp, mutated probes; FP, free probe. (C) ChIP-qPCR of the chsA, csmB, chsC, and chsG genes in the wild-type and CrzA::GFP strains when grown for 24 h in MM and then exposed to increasing concentrations of caspofungin for 1 h. Standard deviations present the average from two independent biological experiments (with 2 technical repetitions each). Statistical analysis was performed using a one-tailed, paired t test compared to the control condition (*, P < 0.05; **, P < 0.005; ***, P < 0.0005).