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Fig. 9. Localization of endogenous Panx1 and Panx3 in normal human skin and mouse spleen. (A-F) Representative images of immunofluorescent labeling for Cx43, Panx1 and Panx3 (red) in normal human facial skin samples and counterstained with the Hoechst nuclear stain (blue). Labeling with either an antibody raised against mouse (A) or human (C) Panx1 identified Panx1 in a focal pattern distributed throughout the epidermis, but most abundantly in the layers of the stratum granulosum and spinosum. (D)Peptide competition with the human PANX1 antibody revealed the loss of specific labeling. (B) Cx43, localized for comparison, was present focally within the upper layers of the epidermis. (E,F) Immunolabeled Panx3 displayed a diffuse cellular distribution within the epidermis (E) that was eliminated by peptide competition (F). Broken lines denote the extent of the cornified layer; dotted lines indicate the edge of the epidermal vital layer. (G,H) Panx1 was localized in 3-week-old normal mouse spleen (G, red), whereas this staining pattern was eliminated after pre-adsorption with cognate peptide (H). Insert in H shows hematoxylin and eosin staining of a parallel spleen section. Bars, 20 µm.
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