Quantification of Bronchial Vascular Biomarkers

Immunohistochemistry was used to detect bronchial vascular biomarkers. The primary monoclonal antibodies against mouse transcription factor (ERG, 1:80), epidermal growth factor (EGF, 1:100), insulin-like growth factor (IGF-1, 1:400), endothelin-1 (1:600), angiogenin (1:400) and amphiregulin (1:500) were purchased from Abcam (Hong Kong, China). The primary monoclonal antibody against mouse Von Willebrand factor (vWF, 1:50) was purchased from CHEMICON International, Inc. MA 01730. The PAP (peroxidase anti-peroxidase) technique was used as previously described [13 (link)-15 (link)]. Positively staining cells were detected using the glucose oxidase-DAB-nickel method [19 (link)]. At least 8 random high-power fields (200 × total magnification) of each mice in the 3 groups should be independently analysed by 2 observers in a blinded fashion on a Leica DM6000B microscope (Leica, Wetzlar, Germany) connected with a Leica Application Suite Version 3.6. The data were expressed as percentages of positive stained areas per unit area of entire lung sections.

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Yao X., Wang W., Li Y., Huang P., Zhang Q., Wang J., Wang W., Lv Z., An Y., Qin J., Corrigan C.J., Huang K., Sun Y, & Ying S. (2015). IL-25 induces airways angiogenesis and expression of multiple angiogenic factors in a murine asthma model. Respiratory Research, 16(1), 39.

Publication 2015

EGF IGF-1 endothelin-1 angiogenin Leica DM6000B microscope

Amphiregulin Angiogenin Biomarkers Bronchial Cells Endothelin 1 Epidermal growth factor Glucose oxidase Igf 1 Immunohistochemistry Insulin like growth factor Lung Mice Microscope Monoclonal antibody Nickel Peroxidase Transcription factor Vascular Von willebrand factor

Corresponding Organization :

Other organizations : Capital Medical University, Beijing Chao-Yang Hospital, Beijing University of Chinese Medicine, King's College London

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Variable analysis

independent variables

Primary monoclonal antibodies against mouse transcription factor (ERG, 1:80), epidermal growth factor (EGF, 1:100), insulin-like growth factor (IGF-1, 1:400), endothelin-1 (1:600), angiogenin (1:400) and amphiregulin (1:500) from Abcam
Primary monoclonal antibody against mouse Von Willebrand factor (vWF, 1:50) from CHEMICON International, Inc. MA 01730

dependent variables

Positively staining cells detected using the glucose oxidase-DAB-nickel method

control variables

PAP (peroxidase anti-peroxidase) technique used as previously described [13-15]
At least 8 random high-power fields (200 × total magnification) of each mice in the 3 groups should be independently analysed by 2 observers in a blinded fashion on a Leica DM6000B microscope (Leica, Wetzlar, Germany) connected with a Leica Application Suite Version 3.6

controls

Positive control: Not specified
Negative control: Not specified

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