br In Vivo Angiogenesis Study br
In Vivo Angiogenesis Study
3 to 4-wk-old BALB/c-nude mice (Slaccas Shanghai Laboratory Animal Co., Ltd., Shanghai, China) were maintained in a pathogen-free environment (23 ± 2 C, 55 ± 5% humidity) on a 12 h light/12 h dark cycle with food and water supplied adlibitum throughout the experimental period. Mice were divided into three groups: control group (injected subcutaneously with 600 ml matrigel containing 200 ml saline), model group (600 ml matrigel containing 200 ml MDA-MB-231or MDA-MB-468 cell suspension) and wogonoside treated group, which were treated with wogonoside (80 mg/kg, gavage, everyday) for ten days. Mice were killed and the gel plugs were excised, photographed, viewed whole-mount of CD31 staining and homogenized in 1 ml PBS buffer, centrifuged to test the content of hemoglobin in the supernatant was by Drabkin’s reagent (Sigma-Aldrich).
Preparation of Cytosolic and Nuclear Extracts
MDA-MB-231 or MDA-MB-468 cells were treated with various concentrations of wogonoside (0, 25, 50 and 100 mM) for 24 h. Nuclear and cytosolic protein extracts were prepared according to the user guide of Nuclear and Cytoplasmic Protein Extraction Kit.
Cells were grown on coverslips and pretreated with wogonoside (100 mM) for 24 h, fixed with 4% paraformaldehyde, permeabilized in 0.2% Triton X-100 and incubated with 3% BSA. After incubated with primary ATPγS tetralithium salt (1:100), cells were exposed to secondary antibodies (1:1000) and stained with DAPI. Cells were observed and photographed with a confocal laser scanning microscope (Fluoview FV 1000, Olympus, Tokyo, Japan).
QUANTIFICATION AND STATISTICAL ANALYSIS
The data shown in the study were obtained from at least three independent experiments and all data in different experimental groups were expressed as the mean± standard deviation (SD). Statistical analyses were performed using a One-Way ANOVA, with post-hoc analysis. Details of each statistical analysis are provided in the figure legends. Differences with P values < 0.05 were considered statistically significant.
DATA AND SOFTWARE AVAILABILITY
The code written for and used in second law of thermodynamics (entropy) study is available from the Lead Contact (Feixiong Cheng, Email: [email protected]) upon reasonable request. Additional data supporting the findings of this study are available within the supplemental information files or from the Lead Contact upon reasonable request.
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A targeted proteomics approach reveals a serum protein signature as diagnostic biomarker for resectable gastric cancer
Qiujin Shen a, Karol Polom b,g, Coralie Williams c, Felipe Marques Souza de Oliveira a, Mariana Guergova-Kuras c, Frederique Lisacek d,e, Niclas G. Karlsson f, Franco Roviello b, Masood Kamali-Moghaddam a,
a Department of Immunology, Genetics and Pathology, Science for Life Laboratory, Uppsala University, Sweden
b Department of General Surgery and Surgical Oncology, University of Siena, Italy
c Ariana Pharmaceuticals, France
d Proteome Informatics Group, SIB Swiss Institute of Bioinformatics, Geneva, Switzerland
e Computer Science Department and Section of Biology, University of Geneva, Switzerland
f Department of Medical Biochemistry and Cell Biology, Institute of Biomedicine, Sahlgrenska Academy, University of Gothenburg, Sweden
g Department of Surgical Oncology, Gdansk Medical University, Gdansk, Poland
Background: Gastric cancer (GC) is the third leading cause of cancer death. Early detection is a key factor to reduce its mortality.