Journal: Frontiers in Immunology

Article Title: Crucial Involvement of IL-6 in Thrombus Resolution in Mice via Macrophage Recruitment and the Induction of Proteolytic Enzymes

doi: 10.3389/fimmu.2019.03150

Figure Lengend Snippet: Inferior vena cava (IVC) ligation-induced deep vein thrombus formation in wild-type (WT) and Il6 −/− mice. (A) Macroscopic appearance of venous thrombi in WT and Il6 −/− mice at 5 and 10 days after IVC ligation. Representative results from six independent animals are shown here. (B) Thrombus mass of WT and Il6 −/− mice at the indicated time intervals after IVC ligation. All values represent the mean ± SEM ( n = 6). ** p < 0.01, WT vs. Il6 −/− . (C) Histopathological analyses of venous thrombi obtained from WT and Il6 −/− mice at 5, 10, and 14 days after IVC ligation. Venous thrombi were stained with hematoxylin and eosin (H&E) or Masson trichrome solution (Masson). Representative results from six independent experiments are shown here (original magnification, ×100). (D) Immunohistochemical detection of Col1A2 proteins in the thrombi obtained from WT and Il6 −/− mice at 5, 10, and 14 days after IVC ligation. (E) Intrathrombotic Col1 gene expression in WT and Il6 −/− mice at the indicated time intervals after IVC ligation. All values represent the mean ± SEM ( n = 6). * p < 0.05, WT vs. Il6 −/− . (F) Laser Doppler analysis of thrombosed blood flow. All values represent the values mean ± SEM ( n = 6 animals). ** p < 0.01, WT vs. Il6 −/− .

Article Snippet: The following mAbs and polyclonal Abs (pAbs) were used for immunohistochemical and double-color immunofluorescence analyses: rat anti-mouse F4/80 mAb (Dainippon Pharmaceutical Company, Japan), mouse anti-IL-6 mAb (Santa Cruz Biotechnology, Dallas, Texas), rabbit anti-mouse IL-6 pAbs, rabbit anti-mouse CCL2 pAbs (Novus Biologicals, Centennial, CO), goat anti-mouse MMP-2 pAbs, goat anti-mouse MMP-9 pAbs (Santa Cruz Biotechnology, Dallas, Texas), rabbit anti-mouse PLAU (uPA) pAbs, rabbit anti-mouse tPA pAbs, rabbit anti-mouse PAI-1 pAbs (Santa Cruz Biotechnology, Dallas, TX), mouse anti-mouse Col1A2 mAb (Santa Cruz Biotechnology), rabbit anti-mouse myeloperoxidase (MPO) pAbs (Neomarkers, Fremont, CA), rabbit anti-mouse CD3 mAb (Abcam, Tokyo, Japan), cyanine dye 3-conjugated donkey anti-rat IgG pAbs, cyanine dye 3-conjugated donkey anti-goat IgG, fluorescein isothiocyanate (FITC)-conjugated donkey anti-rat IgG pAbs, and FITC-conjugated donkey anti-rabbit IgG pAbs (Jackson ImmunoResearch Laboratories, West Grove, PA).

Techniques: Ligation, Staining, Immunohistochemical staining, Gene Expression

Journal: Food & function

Article Title: Suppression of colonic oxidative stress caused by chronic ethanol administration and attenuation of ethanol-induced colitis and gut leakiness by oral administration of sesaminol in mice.

doi: 10.1039/d1fo04120g

Figure Lengend Snippet: Fig. 7 Effects of oral sesaminol administration on the levels of ethanol- induced oxidative-stress proteins (iNOS, CYP2E1, HO-1) and transcrip- tion factor protein (Nrf2) in colonic tissues. (A) WB analyses of iNOS, CYP2E1, HO-1, and β-actin (from top to bottom) in colonic tissue extracts of the mice of groups C, S, E, and ES. Lanes 1–3, group C; lanes 4–6, group S; lanes 7–9, group E; and lanes 10–12, group ES. (B) Protein expression levels of iNOS, CYP2E1, and HO-1 were analysed by WB. The band intensities were measured by densitometry and normalised to β-actin. The relative band intensities (-fold) are shown with the intensity obtained with a subject of group C taken to be 1.0. The data are expressed as means ± SD (n = 5). (C) Nrf2 activation levels were deter- mined by ELISA in colonic tissue extracts, whose protein concentrations were identical (1 μg μL−1) among the four groups. The data are expressed as means ± SD (n = 5). *p < 0.05, **p < 0.01 versus group C, †p < 0.05, ††p < 0.01 versus group S, #p < 0.05 versus group E as assessed by ANOVA with the Tukey–Kramer test.

Article Snippet: The membranes were then incubated with a recommended dilution of one of the following primary rabbit polyclonal antibodies: anti-mouse CYP2E1, anti-mouse inducible nitric oxide synthase (iNOS), anti-mouse zonula occludens-1 protein (ZO-1), anti-mouse occludin, antimouse claudin-1 (each from Abcam), anti-mouse heme oxygenase-1 (HO-1) (Proteintech Group), and anti-mouse β-actin (Genetex; Irvine, CA, USA).

Techniques: Expressing, Activation Assay, Enzyme-linked Immunosorbent Assay