Journal: The Journal of Experimental Medicine

Article Title: Elevated levels of placental growth factor represent an adaptive host response in sepsis

doi: 10.1084/jem.20080398

Figure Lengend Snippet: Effect of PlGF deficiency on tissue mRNA/protein levels of inflammatory and hemostatic markers in a mouse model of endotoxemia. PLGF +/+ (WT) or PLGF −/− (KO) male mice were injected i.p. with or without 16 mg/kg LPS. (A) Shown are the results of quantitative real-time PCR analyses (mRNA copy number per 10 6 copies of 18S) of ICAM-1, VCAM-1, E-selectin, P-selectin, COX-2, and PAI-1 in the heart, lung and liver at 24 h. Data are expressed as means + SD of three independent experiments. *, P < 0.05; **, P < 0.01; and ***, P < 0.0001 compared with untreated controls (and where indicated between PlGF-deficient and wild-type mice). (B) Double immunofluorescence staining for activation markers and CD31 in the liver of wild-type mice treated in the absence (WT) or presence of 16 mg/kg LPS (WT/L) and PlGF −/− mice treated with 16 mg/kg LPS (PKO/L) at 24 h. (a) ICAM-1 (green) and CD31 (red). (b) VCAM-1 (green) and CD31 (red). (c) E-selectin (green) and CD31 (red). (d) P-selectin (green) and CD31 (red). (e) COX-2 (red) and CD31 (green). (f) PAI-1 (red) and CD31 (green). Bars: 132 μm; (insets) 42 μm.

Article Snippet: Immunohistochemistry was performed using the following primary antibodies: hamster monoclonal anti–mouse ICAM-1 (Serotec), rat monoclonal anti–mouse VCAM-1 antibody (BD Biosciences), rat monoclonal anti–mouse E-selectin antibody (BD Biosciences), rat monoclonal anti–mouse P-selectin antibody (Millipore), rabbit polyclonal anti–mouse COX-2 antibody (Cayman Chemical), and rabbit polyclonal anti–mouse PAI-1 antibody (Innovative Research Inc.).

Techniques: Injection, Real-time Polymerase Chain Reaction, Double Immunofluorescence Staining, Activation Assay

Journal:

Article Title: The effect of proteasome inhibitor MG132 on experimental inflammatory bowel disease

doi: 10.1111/j.1365-2249.2008.03872.x

Figure Lengend Snippet: Immunohistochemical staining of phospho-nuclear factor-kappa B (NF-κB) p65 and Ki-67 in interleukin-10-deficient (IL-10−/−) mice treated with MG132. (a) Immunohistochemical staining with anti-phospho-NF-κB p65 antibody. (i) Vehicle-treated IL-10−/− mice; (ii) MG132-treated (15·0 µmol/kg) IL-10−/− mice (original magnification 200×). (b) Immunohistochemical staining with anti-Ki-67 antibody (i, ii) and the Ki-67 labelling index (iii). (i) Vehicle-treated IL-10−/− mice; (ii) MG132-treated (15·0 µmol/kg) IL-10−/− mice (original magnification 200×). The Ki-67 labelling index was defined as the percentage of Ki-67-positive cells per crypt. Six crypts per mouse were evaluated (n = 3 in each group). **P< 0·01 compared with vehicle-treated IL-10−/− mice.

Article Snippet: The dilutions in PBS with 1% BSA were prepared with anti-mouse phospho-NF-κB p65 polyclonal antibody (1:50; Cell Signaling Technology, Inc., Danvers, MA, USA) and anti-mouse Ki-67 monoclonal antibody (1:50; Dako, Copenhagen, Denmark).

Techniques: Immunohistochemical staining, Staining

Journal:

Article Title: The effect of proteasome inhibitor MG132 on experimental inflammatory bowel disease

doi: 10.1111/j.1365-2249.2008.03872.x

Figure Lengend Snippet: The effects of MG132 on histological changes, tumour necrosis factor (TNF)-α gene expression and immunohistochemical staining of phospho-nuclear factor-kappa B (NF-κB) p65 in the colonic tissues of mice with dextran sulphate sodium (DSS)-induced colitis. (a) Histological findings of the rectum at necropsy (on day 10) stained with haematoxylin and eosin. (i) Vehicle-treated mice not administered DSS, (ii) MG132-treated (15·0 µmol/kg) mice not administered DSS, (iii) vehicle-treated mice with DSS-induced colitis and (iv) MG132-treated (15·0 µmol/kg) mice with DSS-induced colitis (original magnification 100×). (b) Histological scores of MG132-treated (15·0 µmol/kg) mice with DSS-induced colitis. The total colitis score is the sum of the three subscores (inflammation severity, inflammation extent, crypt damage score), which was quantified by the scoring system described by Williams et al. (n = 8 in each group). **P < 0·01 between vehicle-treated and MG132-treated mice with DSS-induced colitis. †P < 0·05 compared with respective normal mice not administered DSS. (c) TNF-α gene expression in the colonic tissues of MG132-treated (15·0 µmol/kg) mice with DSS-induced colitis (n = 8 in each group). *P < 0·05 between vehicle-treated and MG132-treated mice not administered DSS. **P < 0·01 between vehicle-treated and MG132-treated mice with DSS-induced colitis. (d) Immunohistochemical staining with anti-phospho-NF-κB p65 antibody. (i) vehicle-treated mice not administered DSS, (ii) MG132-treated (15·0 µmol/kg) mice not administered DSS, (iii) vehicle-treated mice with DSS-induced colitis and (iv) MG132-treated (15·0 µmol/kg) mice with DSS-induced colitis (original magnification 200×).

Article Snippet: The dilutions in PBS with 1% BSA were prepared with anti-mouse phospho-NF-κB p65 polyclonal antibody (1:50; Cell Signaling Technology, Inc., Danvers, MA, USA) and anti-mouse Ki-67 monoclonal antibody (1:50; Dako, Copenhagen, Denmark).

Techniques: Expressing, Immunohistochemical staining, Staining