Fig. 7

Detection of Pka1-regulated proteins in lysates of macrophage-like cells containing C. neoformans. Chromatographic representation of the most abundant peptide and its transition for α-amylase (CNAG_02189) identified from isotopically-labeled peptide or natural peptide for each of the following samples: a macrophage lysate challenged with WT cells grown in 0.2 % glucose; (b) macrophage lysate challenged with WT cells grown in 0.2 % galactose; (c) macrophage lysate challenged with P _GAL7 ::PKA1 cells grown in 0.2 % glucose; (d) macrophage lysate challenged with P _GAL7 ::PKA1 cells grown in galactose 0.2 %. Chromatographic representation of the most abundant peptide and its transition for glyoxal oxidase (CNAG_00407) identified from isotopically-labeled peptide or natural peptide for each of the following samples: (e) macrophage lysate challenged with WT cells grown in 0.2 % glucose; (f) macrophage lysate challenged with WT cells grown in 0.2 % galactose; (g) macrophage lysate challenged with P _GAL7 ::PKA1 cells grown in 0.2 % glucose; (h) macrophage lysate challenged with P _GAL7 ::PKA1 cells grown in 0.2 % galactose. Black indicates isotopically-labeled peptide; red indicates natural peptide. i Quantification of α-amylase identified in the macrophage lysates was based on the area under the curve for the isotopically-labeled peptide versus the natural peptide in WT and P _GAL7 ::PKA1 strains under Pka1-repressed (d) and Pka1-induced (g) conditions. The avergae (± S.D.) amount of peptide present in the sample is reported. j Quantification of glyoxal oxidase identified in the macrophage lysates was based on the area under the curve for the isotopically-labeled peptide versus the natural peptide in WT and P _GAL7 ::PKA1 strains under Pka1-repressed (D) and Pka1-induced (G) conditions. The avergae (± S.D.) amount of peptide present in the sample is reported. Five micrograms of total protein were used for the assays and all assays were performed in triplicate