Skip to main content
. 2008 Dec 29;29(6):1575–1591. doi: 10.1128/MCB.01300-08

FIG. 6.

FIG. 6.

Absence of collagen VI reduces systemic and adipose tissue-specific inflammation. (A) F4/80 immunohistochemical analysis of epididymal fat tissue sections from col6KOob/ob and ob/ob mice, as an indicator of the degree of macrophage infiltration of the tissue. (B) Quantitative real-time PCR analysis for the inflammatory cytokines F4/80, MCP-1, and SAA3 in epididymal and mesenteric fat. Expression levels of all genes were normalized to 18S rRNA (means ± standard errors; n = four mice/group). (C) qRT-PCR analysis of PPAR-γ expression in mesenteric fat. Expression levels were normalized to 18S rRNA (means ± standard errors; n = four mice/group). (D) Phosphorylation states of ERK, JNK, and p38 MAPK in epididymal adipose tissue from col6KOob/ob and ob/ob mice were determined by Western blotting with specific antibodies for their phosphorylated forms and for total levels. Band intensities were quantitated (means ± standard errors; n = three mice/group). (E) LPS challenge in collagen VI-null and WT mice was performed. Circulating IL-6 and MCP-1 levels were measured (upper panel). mRNA expression levels of inflammatory cytokines IL-6 and tumor necrosis factor alpha (TNFα) was measured in epididymal fat by qRT-PCR and normalized to 18S rRNA 90 min after injection of LPS (lower panel) (means ± standard errors; n = four mice/group). (F) CLS were visualized in F4/80-stained epididymal adipose tissue sections. The single asterisk marks a representative CLS, while the double asterisk indicates a remnant lipid droplet after cell death. (G) Apoptotic adipocytes were visualized by TUNEL staining of epididymal tissue sections of col6KOob/ob and ob/ob mice. Brown staining is indicative of a TUNEL-positive nucleus (arrow), while blue staining (hematoxylin stain) represents a nonreactive nucleus (arrowhead). (H) Endoplasmic reticulum stress was measured by PCR analysis for Xbp-1. Primers spanning the splice junction were used to amplify unspliced and spliced forms of Xbp-1 and were separate on a 3% agarose gel. Unspliced Xbp-1 (Xbp1u) is 171 bp, while the spliced Xbp-1 (Xbp1s) is 145 bp and is a positive indication of endoplasmic reticulum stress. All experiments in this figure were performed with 10-week-old col6KOob/ob and ob/ob mice. *, P < 0.05 (data in panels B, C, D, and F were analyzed by Student's t test; data in panel E were analyzed by two-way ANOVA).