Data from: Mucus-derived glycans are inhibitory signals for Salmonella Typhimurium SPI-1-mediated invasion
Data files
Oct 09, 2025 version files 511.34 KB
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Figure_1.xlsx
10.29 KB
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Figure_2.xlsx
243.17 KB
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Figure_3.xlsx
217.04 KB
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Figure_4.xlsx
11.04 KB
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Figure_5.xlsx
15.69 KB
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Figure_6.xlsx
10.61 KB
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README.md
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Abstract
Mucus forms a critical barrier against enteric pathogens like Salmonella enterica serovar Typhimurium. While in vivo studies indicate that secreted, gel-forming mucins and specifically Core 3 glycosylation are protective against S. Typhimurium, the molecular mechanisms involved remain unclear. Here, we demonstrate that native intestinal mucins inhibit Salmonella invasion of colonic epithelial cells by downregulating the type 3 secretion system (T3SS) through suppression of the key virulence regulator, HilD. By measuring changes in the transcriptional profile (qPCR, RNA-seq), proteomics (DIA-based), growth, and HT-29 host cell invasion, our study identifies mucin glycans and specific mucin sugars, namely N-acetyl galactosamine (GalNAc) and N-acetyl glucosamine (GlcNAc), as the components responsible for mucin’s anti-virulence effect, likely via interaction with HilD's putative carbohydrate-binding domain. Notably, we find the native presentation of these sugars is important for activity. These insights provide a mechanistic foundation for mucin-based strategies to combat enteric infections and, given the prevalence of homologous AraC-type regulators in other pathogens, suggest mucins' potential as broad-spectrum anti-virulence agents.
Dataset DOI: 10.5061/dryad.2bvq83c38
Description of the data and file structure
These are Excel files containing the numerical data used to generate the final figures in the manuscript. Specific experimental efforts represented in this dataset include Salmonella invasion assays, qPCR for SPI-1 gene expression, enumeration of colony-forming units to monitor Salmonella growth, and DIA-based proteomics.
Files and variables
File: Figure_1.xlsx
Description: Invasion assays. Log2 fold change in invaded Salmonella cells after 1 h treatment with MUC2 (tab: 1B) or MUC5AC (tab: 1C) relative to medium alone. Columns, Strain ID.
File: Figure_2.xlsx
Description: qPCR data. Log2(fold change in gene expression with MUC2 relative to medium alone) (tab: 2A). RNA-seq data underlying the volcano plot. For each transcript, -log10 False discovery rate, Log2(fold change in gene expression) with MUC2 vs. Medium alone (tab: 2B). qPCR data. Log2(fold change in gene expression with MUC5AC relative to medium alone) (tab: 2C). Growth curve data. The number of colony-forming units at each time point with or without MUC2 (tab: 2D). Proteomics data underlying the volcano plot. For each protein, -log10 p-value, Log2(fold change in expression) with MUC2 vs. Medium alone (tab: 2e).
File: Figure_3.xlsx
Description: Invasion assays. Log2 fold change in invaded Salmonella cells after 1 h treatment with 0.05% MUC2 glycans (tab: 3B) Columns, Strain ID. qPCR data. Log2(fold change in gene expression with 0.05% MUC2 glycans or 0.1% Monosaccharides relative to medium alone) (tab: 3C). RNA-seq data underlying the volcano plot. For each transcript, -log10 False discovery rate, Log2(fold change in gene expression) with 0.1% MUC2 glycans vs. Medium alone (tab: 3C).
File: Figure_4.xlsx
Description: qPCR data. Log2(fold change in hilA expression with MUC2 relative to medium alone), Columns=Mutant ID (tab: 4B). qPCR data. Log2(fold change in gene expression with GalNAc or GlcNAc monosaccharides relative to medium alone) in two different strain backgrounds, WT of HilDQ39E,N44D,H95L (tab: 4E).
File: Figure_5.xlsx
Description: RNA-seq data underlying the heat map. Log2(fold change in gene expression relative to medium alone) Rows = gene name, Column = treatment (i.e., MUC2, glycans, Monsaccharide pool, individual monosaccharides) (tab: 5A). qPCR data. Log2(fold change in hilA expression relative to medium alone), Columns=Treatment type & strain background (tab: 5C). qPCR data. Log2(fold change in prgH expression relative to medium alone), Columns=Treatment type & strain background (tab: 5D). Invasion assays. Number of colony forming units of invaded Salmonella cells after 1 h treatment with medium alone or glycopolypeptides. Rows = Strain (WT, ∆hilD, or HilD-mutant), Column = treatment (tab: 5E)
File: Figure_6.xlsx
Description: qPCR data. Log2(fold change in hilA expression relative to medium alone), Columns=strain background, Row = treatment type (tab: 6B). qPCR data. Log2(fold change in prgH expression relative to medium alone), Columns=strain background, Row = treatment type (tab: 6C)
Access information
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Data was derived from the following sources:
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