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. 2013 Apr 12;8(4):e61380. doi: 10.1371/journal.pone.0061380

Figure 3. SMS1-KO WAT is severely damaged by oxidative stress.

Figure 3

(A) Proteins were extracted from epiWAT and liver of 10-week-old wild-type (WT, n = 3) or SMS1-KO (KO, n = 3) mice and carbonylated proteins were detected by immunoblot analysis using anti-DNP antibody. (B) Samples in (A) were subjected to immunoblot analysis using anti-4-HNE antibody to detect protein modification by ROS. Hsc70 staining served as a standard. (C) Carbonylated proteins in epiWAT of 20-week-old mice were immunohistochemically detected using an anti-DNP antibody (Left panel), and DNP signal intensity in the plasma membrane area was quantified (WT, n = 3; KO, n = 3) (Right panel). (D) mRNAs were extracted from WAT of 10-week-old mice. mRNA expression levels of genes encoding apoptotic factors were assessed by quantitative RT-PCR and normalized to β–actin expression. The wild-type group average was set to 1. n = 6–9 samples per group. (E) Activity of caspase-3 was spectrophotometrically assessed. n = 6 samples per group. (F) mRNA expression levels of genes encoding macrophage-related factors were assessed by quantitative RT-PCR. n = 10–12 samples per group.