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Oxygen-dependent regulation of tumor growth and metastasis in human breast cancer xenografts
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After the treatment the chamber was slowly decompressed to 1 bar over a period of approximately 15 minutes. The mice underwent linda van rijn epub software treatment on days 1, 4, 7, 10, 13 and Day 1 starting measurements was set as linda van rijn epub software 8 days after the injection of tumor cells.
For the separate metastatic study, the mice underwent 17 treatments, i. In the latter case it was given immediately prior to each HBOT for optimal effect. Rabbit serum was used as a blocking agent and H2O2 in methanol was used to quench endogenous peroxidases.
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An avidin biotin peroxidase complex was used to bind to the biotinylated secondary antibody. The cross-section of CD31 positive structures was quantified per mm2 of the entire tumor. The blood vessel diameter was also measured. Also, for standard histologic examination of the tumor specimens, frozen sections from both Linda van rijn epub software tumors and controls were stained with Haematoxylin-Eosin.
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Sections were immediately washed in ddH2O for 3 min. Ki67 positive cells were counted as positive cells per mm2.
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Immunofluorescence IF staining of tissue sections IF staining was performed on cryosections of tissue samples from mice treated with or without HBO for 24 days 5 mice from each group.
The sections were incubated with the secondary antibody for 1 h at room temperature. Sections stained with secondary antibody only were used as negative controls.
Afterwards, the mounted sections were observed under a Zeiss Axioscope fluorescence microscope and photographed using a digital AxioCam mRM camera Zeiss.
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