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. 2021 Aug 4;19(8):e3001360. doi: 10.1371/journal.pbio.3001360

Fig 5. COH1 inactivates the COH2-mediated induction of cuticle-degrading genes during hemocoel colonization.

Fig 5

(A) Quantitative reverse transcription PCR (qRT-qPCR) analysis of the expression of cuticle-degrading genes during cuticle penetration in the mutant ΔCoh2 relative to the wild-type (WT) strain. The chitinase MAA_10456 and the proteases MAA_10199 and MAA_10350 are used as representatives. (B) Chromatin immunoprecipitation quantitative PCR (ChIP-qPCR) analysis shows the occupancy of the COH2::FLAG protein in the promoters of the chitinase and protease genes in the strain WT-COH2-FLAG relative to occupancy in the control strain WT-FLAG, which is set to 1. The fungal strains were grown in the cuticle medium. (C) qRT-PCR analysis of the expression levels of the 3 genes during surrogate hemocoel colonization relative to the expression level during cuticle penetration, which is set to 1. (D) ChIP-qPCR analysis of the occupancy of COH2 in the promoters of the 3 genes during surrogate hemocoel colonization (Hemolymph) relative to the growth in the cuticle medium (Cuticle medium), which is set to 1. The strain WT-COH2-FLAG was used. (E) qRT-PCR analysis of the expression levels of the 3 genes during surrogate hemocoel colonization in the mutant ΔCoh1 relative to the expression level in the WT strain, which is set to 1. (F) ChIP-qPCR analysis of the occupancy of the COH2::FLAG protein in the promoters of the 3 genes during surrogate hemocoel colonization in the strain ΔCoh1-COH2-FLAG (the mutant ΔCoh1 with COH2::FLAG expressed) relative to occupancy in the strain WT-COH2-FLAG, which is set to 1. (G) GFP signal in the fungal cells of 2 strains (WT-PMAA_10199-GFP and ΔCoh1-PMAA_10199-GFP) with the gfp gene driven by the promoter of the protease gene MAA_10199. Top panel: WT-PMAA_10199-GFP on the cuticle (Cuticle); middle panel: WT-PMAA_10199-GFP in the real hemocoel of G. mellonella larva infected by a fungal strain (Hemocoel); bottom panel: ΔCoh1-PMAA_10199-GFP in the real hemocoel. F, fungal cells; H, hemocyte; MGV, mean gray value. Scare bar, 10 μm. The data underlying all the graphs shown in the figure can be found in S1 Data.