Effect of restoration of macrophage A2bAR on liver.

<p>Liver was collected from WT, A2bAR KO, and CD68-Tg mice after 16 weeks of HFD as described in the methods. Relative mRNA expression was determined using the ΔΔCT method with normalization to 18s rRNA. <b>A</b>. mRNA expression of TNF-α in liver. A2bAR KO (n = 5) vs WT (n = 7) p-value = 0.0271; A2bAR KO vs CD68-Tg (n = 6) p-value = 0.0210. <b>B</b>. mRNA expression of IL-6 in liver. A2bAR KO (n = 8) vs WT (n = 7) p-value = 0.0159; A2bAR KO vs CD68-Tg (n = 8) p-value = 0.0080. <b>C</b>. mRNA expression of IRS-2 in liver. CD68-Tg (n = 7) vs A2bAR KO (n = 8) p-value = 0.0300. <b>D</b>. Western blot analysis of liver; one representative (of 3 sets) WT, CD68-Tg and A2bAR KO group shown at 15 minutes post-insulin injection, following 16 weeks of HFD. Levels of mature SREBP-1 (68 kDa), and IRS-2 (185 kDa), were probed by Western blot analysis, using β-actin (43 kDa) as loading control. Quantification of Western Blot results was performed with Image J software (<a href="http://rsb.info.nih.gov/ij/" target="_blank">http://rsb.info.nih.gov/ij/</a>) with normalization to β-actin. WT to A2bAR KO: IRS-2 p-value = 0.0092, SREBP-1 p-value = 0.0154; CD68-Tg to A2bAR KO: IRS-2 p-value = 0.0247, SREBP-1 p-value = 0.0170. Data are averages ± SD. *Student two-tail t-test assuming equal variance was found significant only when p-value <0.05.