Increased TMPRSS2 Expression in Tongue Tissue of Females and Alcohol Drinkers
In the present study, the researchers used immunoblot analysis and immunohistochemistry techniques to determine ACE-2 and TMPRSS2 expression in the human and murine tongue tissue.
Tongue, submandibular glands, and kidneys were acquired from euthanized five-week-old male mice. Half the tissues were fixed in phosphate-buffered formalin for immunohistochemical and histological analysis, while the other half was frozen at -80 °C for the immunoblot assay.
With informed consent, human tongue samples were acquired from patients with primary tongue cancer who underwent surgery. The non-tumorous regions of the tongue were fixed in formalin and embedded in paraffin for the immunohistochemical tests. Three fresh human tongue samples were used for the immunoblot analysis.
Paraffin-fixed tissues were sectioned and mounted on glass slides. Immunostaining was performed using ACE-2 and TMPRSS2 antibodies. Tissues were homogenized and lysed using a lysis buffer after sonication, for the immunoblot analysis. Protein concentration was determined using a protein assay kit and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The immunoblots were incubated with ACE-2 and TMPRSS2 secondary antibodies conjugated with horseradish-peroxidase.
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