Expression of 11β-HSD1, C/EBPα, β, δ and CHOP in adipose tissue of HF diet-fed mice compared to control diet-fed mice.

<p>(A) Real-time PCR measurements of levels of mRNA encoding 11β-HSD1, C/EBPα, β, δ, and CHOP in adipose tissue of HF-fed (black bars) and control diet-fed (white bars) mice. Levels of mRNA, normalized to TBP, are expressed relative to levels in control diet-fed mice, arbitrarily set to 100%. (B-E) Representative western blots (50 µg protein/lane) and quantification of blots probed with; (B) C/EBPα antibody showing levels of the 42 kDa (p42) and 30 kDa (p30) isoforms of C/EBPα as well as total C/EBPα (p42+p30) levels (the intermediate immunoreactive protein indicated by * is likely to be a proteolytic product of p42); (C) C/EBPβ antibody showing levels of LAP (38 kDa LAP* +35 kDa LAP) and the 20 kDa LIP isoforms of C/EBPβ as well as total C/EBPβ (LAP+LIP) levels (inset shows LIP:LAP ratio); (D) C/EBPδ antibody and (E) CHOP antibody showing levels of respective proteins in adipose tissue of mice fed HF diet (HF, black bars) or control diet (C, white bars). Blots were stripped and reprobed with β-tubulin antibody, as loading control. In C/EBPβ and CHOP westerns, all samples were analysed in the same gel but not all in adjacent lanes. C/EBP levels were quantified relative to β-tubulin levels using ImageJ analysis and are expressed in arbitrary units (AU). All values are mean±SEM; n = 16/group (A) or 4/group (B-E). *, p≤0.05; **, p<0.001.