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In order to determine the effects of the deletion of CrT on body and brain weights, animals were weighed prior to testing and brains were weighed after behavioral testing.
Brains were weighed and homogenized (10% w/v) in homogenization buffer, 20 mM Tris, 250 mM sucrose, 1 mM EDTA, 1 mM EGTA with freshly prepared 100 mM phenylmethylsulfonyl fluoride, 5 µg/ml pepstatin A and a protease inhibitor cocktail (Complete, Roche Diagnostics, Indianapolis, IN).
The frozen brains were weighed and homogenised (10% w/v) in PBS containing 2% Sarcosyl.
Mice brains were weighed and homogenized in 10× volume of homogenate buffer.
The brains were weighed and stored at −70°C prior to AVT and IT analyses.
At the end of the experiment, following perfusion (see below), brains were weighed.
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The dissected prefrontal regions from each brain were weighed frozen (∼100 mg) and processed.
Mouse hemi-brains were weighed and homogenized in 10× volume of buffer [50 m m Tris base (pH 8), 274 m m NaCl, 5 m m KCl, with 1× protease and phosphatase inhibitors (Thermo Scientific, Waltham, MA)].
Wet liver and brain were weighed and flash frozen in 2-methylbutane on dry ice and stored at −80 °C until further processing.
Hemispheres of mouse brain were weighed, proteins extracted and calprotectin (S100a8/a9) was quantified as recommended by the ELISA manufacturer (Immundiagnostik, Bensheim, Germany).
The brain was weighed and an image was recorded with Olympus SZX7 microscope with an Infinity 1 camera and then freeze-dried and weighed again before it was homogenized in PBS buffer.
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