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The double stained cells in eight randomly selected areas along the IBZ were measured per section and 3 sections were taken per animal.
The following parameters were measured per section: total number of structures, area of clusters, total number of cells per structure, and number of Ki67 positive cells within those structures.
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The length of 10 well-oriented villi with both side-crypts visible was measured per cross-section, and the mean length was calculated.
Density measure per section was computed by averaging the measurements obtained from the two 500 μm-wide traverses.
At least three sections, separated by 200 μm, were measured per animal.
30 villi were measured per mouse.
To measure cortical thickness, the average from three measurements per section was calculated in the metaphyseal region.
Muscle fiber sizes were measured using the ImageJ software (National Institute of Health, Baltimore, MD) and at least 400 muscle fibers per animal were measured (8 images per section and 5 sections at 50 μm interval per mice).
The areas of SVZ in the ipsi- and contralateral hemispheres were traced and measured in three sections per animal, 10 sections (50 μm) apart, using the software 'Stereo investigator ver. 6' (MBF Bioscience, Williston, VT, USA).
All sections were mounted and stained, with 4 sections per slide, and gland area was measured from one section on each slide collected.
The cell perimeter and sectional area were measured in 100 adipocytes per section (three sections for one mouse and five animals per group), and data analysis performed using the MCID Basic software (Imaging Reseaech Inc., Ontario, Canada) for digital image processing.
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