Fig. 5

PPARG overexpression was insufficient to rescue anti-adipogenic effects of M^(IFNG/LPS) macrophages. a Representative immunoblots of the adipogenesis transcription factors CEBPB, PPARG, and CEBPA in early differentiating ASC (day 3) exposed to macrophage-CM. b Corresponding mRNA levels of day 3 differentiated cells. Data were normalized to the 18S rRNA reference gene and shown as mean –ΔCt, n=3. c Representative immunoblot of PPARG in proliferating ASC 72h after lentiviral transduction. d Immunoblotting of adipocyte marker proteins in PPARG- and puromycin resistance gene transduced ASC differentiated to adipocytes for 14 days in the presence of macrophage-CM. e Flow cytometry analysis of day 14 differentiated PPARG- or puromycin resistance overexpressing ASC. Values depict relative numbers of large lipid containing adipocytes (stained with BODIPY-493/503) compared to Mo-CM-treated cells, n=4. For panel b, statistical significance was determined by one-way ANOVA and Dunnett’s test for multiple comparison, for panel e, p values were calculated by unpaired Student’s t test. Data are shown as mean ± SEM