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Tissue preparation for light microscopic immunohistochemistry/immunofluorescence Male C57Bl/6 J mice (n = 3, 1213 weeks old, weighing 2731 g) were anesthetized with a mixture of ketamine and xylazine as described above, and perfused transcardially with 10 ml PBS (pH 7.4), followed by 40 ml 4% paraformaldehyde in 0.1 M PB (pH 7.4)

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This gradual titration helps minimise gastrointestinal side effects