Journal: bioRxiv
Article Title: Salmonella lipopolysaccharide stimulates uptake of long-chain fatty acids in the small intestine
doi: 10.64898/2026.05.19.726283
Figure Lengend Snippet: (A) qPCR analysis of lipid metabolism mRNAs ( Cd36 , Fabp1 , and Apoa4 ) expression in small intestines of antibiotic-treated mice injected intraperitoneally with 1 µg of pathogen-associated molecular patterns (PAMPs): lipoteichoic acid (LTA), flagellin, or ultra-pure lipopolysaccharide (LPS) (n=6 mice per group). (B) In vivo uptake of C16:0 BODIPY in small intestinal epithelial cells (IECs) from antibiotic-treated mice injected intraperitoneally with the PAMPs shown in (A). Mice were injected daily for 3 days. Mice were gavaged with C16:0 BODIPY 12 h after the final treatment, and IEC fluorescence was quantified by flow cytometry. Analysis was performed on live EpCAM⁺ cells. Representative histograms and quantification of MFI are shown (n=7–8 mice per group). (C) Schematic of S . Typhimurium lipopolysaccharide (LPS) biosynthesis and mutant strains used in (D–F). The LPS biosynthesis pathway generates penta-acylated LPS, which is converted to hexa-acylated LPS by the acyltransferase MsbB (LpxM); the Δ msbB mutant retains penta-acylated LPS. (D) qPCR analysis of lipid metabolism mRNAs ( Cd36 , Fabp1 , and Apoa4 ) in small intestines of germ-free or antibiotic-treated mice infected by intragastric gavage with parental wild-type or mutant S . Typhimurium IR715 strains shown in (C) (n=5–8 mice per group). (E) Mass spectrometry–based lipidomic analysis of IECs from uninfected or wild-type or mutant S . Typhimurium IR715–infected mice is shown as bar plots. Relative abundance of 14:0, 16:0, and 18:0 fatty acid–containing glycerolipids is shown (n = 3–4 mice per group). (F) In vivo uptake of C16:0 BODIPY. Antibiotic-treated mice were infected with parental wild-type or mutant S . Typhimurium IR715 strains and orally gavaged with C16:0 BODIPY. IEC fluorescence was quantified by flow cytometry. Representative histograms and MFI quantification are shown (n=6–11 mice per group). LPS, lipopolysaccharide; PAMPs, pathogen-associated molecular patterns; i.p., intraperitoneal; Abx-treated, antibiotic-treated; qPCR, quantitative real-time PCR; sm. int., small intestine; LTA, lipoteichoic acid; IEC, intestinal epithelial cell; Veh, vehicle; S . Tm, Salmonella Typhimurium; Uninf., uninfected; WT, wild-type. MFI, median fluorescence intensity. Data shown in panels A, B, D, and F are representative of 2–3 independent experiments. Each bar graph data point represents one mouse. Significance was determined by one-way ANOVA.
Article Snippet: For i.p. administration, mice received 40 ng/g body weight of ultrapure flagellin (InvivoGen, cat. #tlrl-epstfla), lipopolysaccharide (InvivoGen, cat. #tlrl-pb5lps), or lipoteichoic acid (InvivoGen, cat. #tlrl-pslta).
Techniques: Expressing, Injection, In Vivo, Fluorescence, Flow Cytometry, Mutagenesis, Infection, Mass Spectrometry, Real-time Polymerase Chain Reaction
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