Journal: Nature microbiology

Article Title: Human gut bacteria produce structurally related monoglycolipids with contrasting immune functions

doi: 10.1038/s41564-025-02141-1

Figure Lengend Snippet: Extended Data 3. In vitro and in vivo assays for NKT cell modulation by BfaGCs. a) Schematic process illustrates the in vitro antigen presentation assay. Bacterial lipid extracts were loaded onto biotinylated CD1d molecules, which were subsequently immobilized on streptavidin-coated plates. After washing, NKT cell hybridomas (24.7) were added and incubated overnight. IL-2 secretion was measured by ELISA to assess NKT cell activation. b) Representative gating strategies for colonic NKT cell analysis by flow cytometry are shown. NKT cell refers to the CD1d tetramer + cells within the CD3 + CD45 + TCRβ + T cell population. c) Representative flow cytometry plots of unloaded CD1d tetramer controls are shown.

Article Snippet: After overnight incubation at 37°C, ligand-loaded CD1d (0.25 ug per well) was bound onto streptavidin-coated plates (R&D Systems #CP003).

Techniques: In Vitro, In Vivo, Immunopeptidomics, Incubation, Enzyme-linked Immunosorbent Assay, Activation Assay, Cell Analysis, Flow Cytometry