Fig 2: Levels of MIF in native HAT and IAT.Adipose tissue from healthy donor sites and acutely inflamed donor sites were homogenized. MIF levels were measured by ELISA. A MIF levels of native HAT and IAT B MIF levels of supernatants derived from HAT and IAT. Data are presented in mean MIF concentration ± SEM. Statistically significant differences are indicated by asterisks (*, P<0.05).

Fig 3: Flow cytometry of labeled ATM from epididymal fat pads from WT and Mif -/- mice.FITC-labeled macrophages were injected retro-orbitally into WT and Mif -/- mice after inducing a local inflammation in the epididiymal fat pads by LPS. After 48 hours, mice were sacrificed and FITC-positive macrophages were measured by flow cytometry. Doublets were excluded by sideward scatter (SSC) and forward scatter (FSC), macrophages were defined as CD45, CD11b, and F4/80 positive cells. The results show a significant lower number of FITC-positive macrophages in Mif -/- mice when compared to WT mice. Data is presented as % of FITC-positive ATMs ± SEM. Statistically significant differences are indicated by asterisks (*, p<0.05).

Fig 4: Correlation of MIF protein with donor’s BMI.MIF protein levels from native adipose tissue were correlated (Spearman’s rank order correlation) with the BMI of the donor. The linear regression curve is depicted by a solid line, the 95% confidence interval by a dotted line. A MIF protein levels and BMI from donors of HAT show a significant positive correlation (p<0.05, Spearman r = 0.39). B MIF protein levels and BMI from donors of IAT show no significant correlation (p = 0.32, Spearman r = -0.23).

Fig 5: In vitro scratch assay of dermal fibroblasts and supernatants from IAT and HAT.Wound healing of human dermal fibroblast monolayers was evaluated by an in vitro scratch assay. Fibroblasts were incubated with supernatants of samples from IAT and HAT, MIF-dependency was evaluated by anti-MIF antibodies. Medium containing 10% FCS served as a PC and medium containing 0% FCS served as a NC. Experiments were performed in the presence (A, B) and absence (C, D) of the proliferation inhibitor Mitomycin C. A Supernatants from IAT show a decreased induction of fibroblast proliferation and migration (no Mitomycin C). B Fibroblast proliferation and migration is increased by addition of MIF-antibodies (no Mitomycin C). C Supernatants from IAT show a decreased induction of fibroblast migration (10 µg/ml Mitomycin C). D Fibroblast migration is increased by addition of MIF-antibodies (10 µg/ml Mitomycin C). All measurements were normalized to the negative control of Fig 7A which was set as 1. Data are presented in relative migration distance of fibroblasts ± SEM. Statistically significant differences are indicated by asterisks (*, p<0.05; **, p<0.01; ***, p<0.001).