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Figures
- Fig. 1.
Inverse pattern of prostasin versus matriptase expression during the course of epidermal differentiation. Human skin tissue sections were immunostained for prostasin (A,B), matriptase (C,D), and HAI-1 (E,F). Other sections were also stained with a non-specific mouse IgG antibody as a negative control (data not shown). Representative examples of the staining observed are presented. Cell nuclei were lightly counterstained with hematoxylin. Scale bars: 100 µm in A, C, and E; 40 µm in B, D, and F.
- Fig. 2.
Expression of prostasin, matriptase, activated matriptase, and HAI-1 in human hair follicles. Cross sections of human hair follicles were immunostained with the prostasin-specific mAb YL11 (A), matriptase-specific mAb M24 (B), the HAI-1-specific mAb M19 (C) or the activated matriptase-specific mAb M69 (D) and counterstained with hematoxylin. Sections were also stained with a non-specific mouse IgG antibody as a negative control (data not shown). Representative examples of the staining observed are presented. Scale bars: 30 µm.
- Fig. 3.
Expression of prostasin, matriptase and HAI-1 in human skin. Human skin tissue sections containing the epidermis, hair follicles, and sebaceous glands were immunostained with the prostasin mAb YL11 (prostasin; A,B), matriptase-specific mAb M24 (matriptase; C,D), or the HAI-1-specific mAb M19 (HAI-1; E,F), and counterstained with hematoxylin. Other sections were also stained with a non-specific mouse IgG antibody as a negative control (data not shown). Representative examples of the staining observed are presented. The different regions of the skin section are as indicated, with the sebaceous glands shown at higher magnification in the lower panels. Scale bars: 100 µm in A, C, and E; 40 µm in B, D, and F.
- Fig. 4.
Prostasin is localized primarily at membrane protrusions in HaCaT human keratinocytes. The subcellular localization of prostasin (A,D; red) in HaCaT human keratinocytes were analyzed by immunofluorescent staining with the prostasin mAb YL11. The cells were also stained for F-actin using Alexa 488-labeled phalloidin (B,E; green) and nuclei using DAPI (blue), as a counterstain. The staining is also presented as merged false-color images (C,F). Scale bars: 20 µm.
- Fig. 5.
Matriptase and HAI-1 are primarily expressed at the cell-cell junctions. The subcellular localizations of matriptase (A; green) and HAI-1 (D; green) in HaCaT human keratinocyte cultures were analyzed by immunofluorescent staining with the matriptase-specific mAb M24 and HAI-1-specific mAb M19. The cells were also stained for F-actin using Alexa 594-labeled phalloidin (B,E; red) and nuclei using DAPI (blue), as a counterstain. The staining is also presented as merged false-color images (C,F). The staining of matriptase at different types of cell-cell contacts is as indicated. Scale bars: 20 µm.
- Fig. 6.
Human keratinocytes retain active prostasin. HaCaT human keratinocytes were transiently exposed to PBS as a non-activation control (lanes 1 in A,B; lanes 1 and 3 in C) or a pH 6.0 buffer to induce zymogen activation of matriptase and prostasin (lanes 2, in A,B; lanes 2, 4, and 5 in C). A separate sample of the lysate used for lane 5 was subjected to immunodepletion of prostasin using the mAb YL11-conjugated Sepharose and analyzed in lane 6 (lane 6 in C). The conditioned buffer following the induction of zymogen activation was collected and concentrated to the volume equal to that of the cell lysate (lane 7). These samples were analyzed for the species of matriptase (A, MTP), HAI-1 (B, HAI-1) and prostasin (C). The identity of the protein bands are as indicated.
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