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 Post subject: One section on each slide was used as a control to assess no
PostPosted: Mon Jan 22, 2018 3:17 am 
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Joined: Wed Jul 31, 2013 1:53 am
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The diameter of marrow microvessels was determined as the average diameters of microvessels in the same area used to measure MVD, using a digitizing image analysis system. To evaluate PPARc positive expression in distal femur, the slides were scanned by LeicaQ550CW image analyzer and the average of absorbance from 30 positive expression Torin 1 mTOR inhibitor cells on each slide was presented as mean 6 SD. Immunohistochemical analysis of bone marrow BMP-2 and BMP-7 expressions were performed following the manufacturers’ instructions: Antigen retrieval was performed by high temperature and pressure treatment of the sections in citrate buffer. After a wash in PBS, sections were incubated with hydrogen peroxide blocking agent to quench endogenous tissue peroxidases and then washed in PBS. The slides were incubated with Ultra V Block for 30 min at room temperature. The primary antibodies were incubated according to the manufacturer’s protocol. Slides with serial sections were incubated with the primary antibodies in a humidified chamber at 4uC overnight, followed by being incubated with streptavidin-perosidase avidin for 10 min at room Trichostatin A HDAC inhibitor temperature. Diaminobenzadine staining was done by incubating the sections with coloring reagent. The sections were counterstained with Harris’ haematoxylin, dehydrated through increasing concentrations of alcohol and mounted with coverslips.


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