Fig. 1: Protein O-GlcNAcylation differs significantly between Y. pestis grown under conditions mimicking its two typical niches.

a Workflow for obtaining bacterial protein samples of Y. pestis grown under various conditions. b The O-GlcNAcylations of the wild-type Y. pestis strain were found to be significantly different under seven different growth conditions. The experiment was independently repeated at least three times, yielding consistent results. Protein samples were obtained using the method described in a, separated on a 12% SDS-PAGE gel, and immunoblotted with a mouse monoclonal anti-O-GlcNAc antibody. Long and short exposures were used to provide comprehensive information. Lanes 1-7 correspond to Fv, Mh, 10 °C in TMH, 42 °C in TMH, 37 °C in pH 6 TMH without calcium, 26 °C in TMH with 2.5% NaCl, and 37 °C in TMH without Ca²⁺ and Fe²⁺. c Technical roadmap for quantitative proteomic analysis of O-GlcNAcylation of protein in Y. pestis grown under the Fv and Mh conditions. d Statistics plot of the proteins and sites with different abundances under the Fv and Mh conditions in triplicates. Proteins with fold change (FC) ≥ 1.2 were considered to be up-regulated. e Statistical plot predicting the subcellular localization of the differentially modified proteins. f Statistical plot showing the distribution of differentially modified proteins in secondary Gene Ontology (GO) annotations. BP biological process, CC cellular component and MF stands for molecular function.