Figure 8.

Primary adipocytes differentiated from GR flox and Adipo GRKO mice showing significantly reduced GR mRNA expression and the effect of GR stimulation on immunoregulatory genes. (A) Representative images of undifferentiated, differentiated lipid laden, and Oil-Red-O-stained lipid droplets in primary GR Flox and Adipo GRKO adipocytes on day 10. Scale bar: 5 μm. (B) Stained lipid droplets were quantified using the extracted Oil-Red-O stain and absorbance was measured at 492 nm. (C) Representative immunofluorescence staining of adiponectin and GR proteins in differentiated GR Flox and Adipo GRKO adipocytes. (D) Adipoq mRNA expression as measured by qPCR in undifferentiated and differentiated GR Flox and Adipo GRKO adipocytes on day 10. (E) GR mRNA expression as measured by qPCR in differentiated GR Flox and Adipo GRKO adipocytes on day 10. (F) Primary GR Flox and Adipo GRKO adipocytes were treated with vehicle (control) or 100 nM dexamethasone (grey bars) for 3 hours. The mRNA gene expression of Cxcr2, Cemip, Coch, Ighm, and Dnase1 was measured by qPCR. All values represent mean ± SD. (D)-(F) Data from 3 independent experiments (in duplicate for each experiment) were combined and statistically analyzed using either 2-way ANOVA with Tukey's multiple comparisons (D) or Student's t-test [E-F). * P < 0.05, ** P < 0.01, ***P < 0.001.

Abbreviations: Adipo GRKO, adipocyte-specific GR knockout; GR, glucocorticoid receptors; GR Flox, C57BL/6J GR flox/flox; qPCR, quantitative PCR.