Fig. 3

M^(IFNG/LPS) CM impairs the formation of mature lipid-laden adipocytes. a Representative images of human ASC differentiated to adipocytes for 14 days in the presence of distinct macrophage-CM. Lipid droplets and nuclei were visualized by BODIPY493/503 (green) and Hoechst 33342 (blue), respectively. Sizebar = 100μm. b Flow cytometry analysis of differentiated adipocytes at day 14. Shown are an overlay of histrograms (left panel) and relative numbers of large lipid containing adipocytes (stained with BODIPY-493/503) compared to Mo-CM-treated cells, n=6. c, d mRNA-expression levels of selected adipokines (c) and angiogenesis- or inflammation associated cytokines (d) in day 14 differentiated adipocytes. Data were normalized to the 18S rRNA reference gene and shown as mean –ΔCt, n=3. e Representative microscopy images of day 14 differentiated adipocytes exposed to CM of pro- and anti-inflammatory CD14⁺ macrophage-CM. f Quantification of large lipid droplet containing cell numbers exposed to CD14⁺ macrophage-CM after 14 days of differentiation, n=4. Asterisks indicate p values <0.05 (*), <0.01 (**), <0.001 (***), and <0.0001 (****). Statistical significance was determined by using one-way ANOVA and Dunnett’s test for multiple comparison. Data are shown as mean ± SEM