FIG 1 .

The MpkA-CWI integrity pathway is activated during the CPE. (A) A. fumigatus CEA17 conidia (10⁴) were inoculated on solid minimal medium (MM) supplemented with glucose and different caspofungin concentrations for 5 days at 37°C. (B) Different concentrations of A. fumigatus CEA17 conidia (left) were inoculated in liquid MM supplemented with glucose and different caspofungin concentrations for 16 h at 37°C. (C) Western blotting assay of MpkA phosphorylation in response to increasing caspofungin concentrations. Anti-p44/42 MpkA or anti-44/42 MpkA antibodies were used to detect the phosphorylation of MpkA and total MpkA, respectively. Anti-γ-tubulin antibody was used as a loading control. Signal intensities were quantified using the ImageJ software, and ratios of (P)-MpkA to MpkA were calculated. A Coomassie brilliant blue (CBB) gel of the protein extract served as an additional loading control. (D) Heat map and hierarchical linkage clustering (as determined by MeV software) of RT-qPCRs of the chitin synthase gene mRNA accumulation in the presence of caspofungin in the wild-type, ΔmpkA, and ΔrlmA strains. Strains were grown for 16 h at 37°C and transferred to increasing caspofungin concentrations for 60 min. The results are expressed as the average of the fold increase of the control cDNA (without caspofungin) for a specific gene of three biological independent experiments (with 2 technical repetitions each; see Fig. S1).