Fig. 3 | Nature Communications

Fig. 3

From: Leptin induces TNFα-dependent inflammation in acquired generalized lipodystrophy and combined Crohn’s disease

Fig. 3

Leptin induces pro-inflammatory immune responses in the AGLCD patient and improves wound healing in vitro. a Leptin serum concentrations of the AGLCD patient before (−) and after (+) 4 days of rLeptin substitution. b–g Flow and mass cytometric analysis of the effects of rLeptin treatment on PBMCs in the AGLCD patient after 4 and/or 7 days. b Lipid droplet content assessed by BODIPY staining. c Heat map displaying the relative changes in mean expression (compared to before rLeptin substitution) of various functional markers in different cell subsets (mass cytometry antibody panel B in Supplementary Table 2) (red, fold change above 2 (increased expression); yellow, fold change = 0.1 (decreased expression)). d Perforin expression measured as mean fluorescence intensity (MFI). e TNFα-expressing cells and the respective MFI. f Two-dimensional projections of single cell data generated by t-SNE of mass cytometry data (antibody panel A in Supplementary Table 2) show the effects on (g) frequencies of different cell subsets. Heat colors of expression levels of selected markers on t-SNE maps have been scaled for each marker individually (red, high expression; blue, low expression). h Serum levels of different immune cell-derived factors measured by CBA ij Oxygen consumption rate (OCR) assessed by Seahorse analyses in monocyte-derived macrophages of a healthy donor differentiated in the presence of serum from the AGLCD patient before (“leptin-free”) and after in vitro or in vivo leptin/rLeptin substitution (performed in at least triplicates, error bars show ± SEM, two-way ANOVA with post-tests and Holm-Sidak correction). The corresponding extracellular acidification rates (ECAR) are reported in Supplementary Fig. 7. k Scratch assay with human T84 intestinal epithelial cells in the presence of leptin to assess in vitro wound healing. lm Effects of leptin on the transepithelial electrical resistance (TEER) across a monolayer of T84 cells after challenge with TNFα and IFNγ to induce leakage (error bars show SD). *P < 0.05, **P < 0.01, ***P < 0.001, two-tailed unpaired t-tests (where applicable). The source data are provided as a Source Data file.

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