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. 2024 Oct 30;10(11):749. doi: 10.3390/jof10110749

Figure 2.

Figure 2

Cell wall integrity pathway and protein kinase A activity are important for ppo-altered susceptibility to CAS. (a) Western blot analysis for wild-type A. fumigatus, ΔppoA, and ΔppoC, grown in liquid MM for 16 h at 37 °C and transferred, or not, to 2 µg/mL of CAS for 0, 10, 30, and 60 min. OEppoA and OEppoC were grown in liquid MM for 16 h at 37 °C. MpkA-P was identified using P-p44/42 antibody, while histone H3 was identified by the anti-histone H3 antibody. (b) PKA activity of the wild-type and ppo mutants. The strains were grown as described in (a) and the samples were processed according to the PKA colorimetric activity kit (https://www.thermofisher.com/order/catalog/product/br/pt/EIAPKA, accessed on 29 September 2024) and normalized using the total protein. The results are the average of three repetitions ± standard deviation. *** p-value < 0.001 (c,d) FunCat analysis of the DEGs that were up- and downregulated in the ΔppoC and OEppoC mutants. (e) LC-HRMS analysis of the supernatant of the wild-type and OEppoC strains grown in liquid MM for 5 days at 37 °C. The results are the average of three repetitions ± standard deviation.