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For the histologic evaluation of skin tissues, the area of the skin lesion was fixed in 10% formalin and then embedded in paraffin. The paraffin-embedded sections (5 μm thickness) were stained with hematoxylin and eosin (H&E) and toluidine blue (Sigma Aldrich Chemical Co. St Louis, MO) for the assessment of inflammatory cells. The total (200X magnification) and degranulated (400X magnification) mast cells per 3 high-power fields were counted and averaged per mouse (total 5 mouse) using ImageJ 1.47 software (NIH, Bethesda, MD)61.
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