Fig. 7: GC-induced macrophage deactivation contributes to adiposity by inhibiting STAT3 signaling in adipocytes.

mRNA levels in ATMs of male (a), female (b) LoxP, and mKlf9KO mice treated with Dex (5 mg/kg) (n = 5 mice) or vehicle (n = 6 mice) for 6 weeks. c STAT3 binding site analysis of indicated genes. STAT3 phosphorylation in beige adipocytes treated with CM from mPMs pretreated with Dex (100 nM) or vehicle for 4 h (d) (without Dex (e)) and then stimulated with IL-4 (20 ng/ml) and LPS (1 ng/ml) for another 12 h. (n = 3 independent experiments). f STAT3 phosphorylation in WAT treated as in (a) (n = 3 mice). g mRNA levels in beige adipocytes treated with CM containing S3I-201 (100 μM) from indicated macrophages pretreated with IL-4 (20 ng/ml) and LPS (1 ng/ml) for 12 h (n = 3 independent experiments). h KLF9 mRNA in hPBMC-MΦ treated with cortisol (1 μM) or vehicle for 16 h (n = 5 independent experiments). i mRNA levels in beige adipocytes treated with CM from hPBMC-MΦ pretreated with LV-KLF9 or LV-GFP for 24 h and then stimulated with IL-4 (20 ng/ml) and LPS (1 ng/ml) for another 12 h (n = 6 independent experiments). j STAT3 phosphorylation in beige adipocytes treated as in (i) (n = 3 independent experiments). k mRNA levels in hPBMC-MΦ pretreated with sh-KLF9 or sh-Con for 24 h and then treated with cortisol (1 μM) or vehicle for 4 h and then co-stimulated with IL-4 (20 ng/ml) and LPS (1 ng/ml) for another 12 h (n = 6 independent experiments). STAT3 phosphorylation (n = 3 independent experiments) (l) and mRNA levels (n = 6 independent experiments) (m) in beige adipocytes treated with CM from hPBMC-MΦ treated as in (k). n mRNA levels in beige adipocytes treated with CM containing S3I-201 from hPBMC-MΦ treated as in (i) (n = 6 independent experiments). Data are represented as mean ± SEM., unpaired two-tailed Student’s t tests were performed in (e, h, i, and j), one-way ANOVAs were performed in (d, f, and l), or two-way ANOVAs were performed in (a, b, g, k, m, and n).