Fig. 3: Energy-metabolic pathways are suppressed by LTβR.

Neutrophils were detected from BM of Ltbr^fl/fl × Mrp8-Cre (LTβR^ΔN) and control littermates by flow cytometry (a, f, g) or by negative magnetic enrichment (STEMCCELL technologies) (b–e) and metabolic parameters were measured. Mitochondrial mass was measured as mean intensity of Mitotracker staining (a) and mean Tom20 immunofluorescence intensity by confocal microscopy (b). c, d Tom20 foci indicative of mitochondria were quantified from z-stacked confocal micrographs of Ly6G+ cells (red) stained with Tom20 antibody (green) and quantified by Imaris. Organization of Tom20 in clusters and the 3D-shape reflected in z-stacks can lead to multiple individual foci recorded for one mitochondrium. Scale bar, 500 nm e Representative TEM images (right) and blinded quantification of granule and mitochondrial content (left). Red arrows indicate individual mitochondria. Scale bar, 500 nm. f Neutrophil mitochondrial superoxides were measured by MitoSox and quantified by flow cytometry as mean fluorescence intensity. g Representative gating and histograms (left) and quantification (right) of neutrophil oxidative burst assay at baseline and in response to fMLP (fMLP N-formylmethionine-leucyl-phenylalanine; DHR dihydrorhodamine-123). h Fluorescent glucose uptake was quantified using the fluorescent glucose analog 2-NBDG (Invitrogen) by flow cytometry. Student’s t test, data from 3–6 mice/group from 2 or more independent experiments. Error bars represent SEM.