Rab11 Is Associated with Epidermal Lamellar Granules

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Rab11 Is Associated with Epidermal Lamellar Granules Akemi Ishida-Yamamoto, Mari Kishibe, Hidetoshi Takahashi, Hajime Iizuka Journal of Investigative Dermatology Volume 127, Issue 9, Pages 2166-2170 (September 2007) DOI: 10.1038/sj.jid.5700850 Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Rab11 is strongly expressed in differentiated epidermal keratinocytes, and is partially colocalized with TGN46. (a–f) Confocal laser scanning microscopy. Lower (a–c) and higher (d–f) magnification views. (a, d) Rab11 (rabbit polyclonal antibody, green) is expressed from the middle spinous layer to the superficial granular layer. Staining is more intense at the apical parts of each cell. (b, e) TGN46 staining (sheep polyclonal antibody, seen in red) is seen around the nuclei in all cells. Merged images are shown in (c and f). There is partial overlap between Rab11 and TGN46 staining. Differential interference microscopy images are superimposed upon immunofluorescent images. (g) Post-embedding immunoelectron microscopy staining of epidermal keratinocytes with Rab11 and TGN46 antibodies. TGN46 labels (10-nm gold particles) are localized on the trans-side of Golgi apparatus, whereas Rab11 labels (5-nm gold particles, arrows) are associated with TGN and other vesicular structures. (a–c) Bar=20μm. (d–f) Bar=10μm. (g) Bar=100nm. Journal of Investigative Dermatology 2007 127, 2166-2170DOI: (10.1038/sj.jid.5700850) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Rab11 expression is overlapped with that of various lamellar granule molecules. Confocal laser scanning microscopy. (a, d, g, j) Rab11 and (b) LEKTI, (e) corneodesmosine, (h) cathepsin D, and (k) glucosylceramides are double stained. Merged images are shown in (c, f, i, and l). Differential interference microscopy images are superimposed on immunofluorescent images. Rab11 and lamellar granule molecules are colocalized in superficial spinous cells and granular cells. Note that Rab11 labels are less intense in (d and j) than in (a and g) probably because a mouse monoclonal Rab11 antibody was used for the former, whereas a rabbit polyclonal Rab11 antibody was used for the latter. Bar=20μm. Journal of Investigative Dermatology 2007 127, 2166-2170DOI: (10.1038/sj.jid.5700850) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Rab11 is associated with LEKTI-positive lamellar granules. Double staining of Rab11 (5-nm gold labels, small arrows) and LEKTI (10-nm gold labels, large arrows) using cryo-ultra microtomy. (a) A lower magnification view of the stratum corneum (C), the most superficial stratum granulosum (G1), the second (G2), and the third stratum granulosum (G3) is shown. The rectangular areas marked in (a and b) are shown at higher magnification in (b and c) respectively. Note that Rab11 labels are widely distributed along the tubular-vesicular structures with partial lamellar internal structures extending from the proximity of the nucleus (Nu) to the cytoplasm. (a) Bar=2μm. (b) Bar=500nm. (c) Bar=100nm. Journal of Investigative Dermatology 2007 127, 2166-2170DOI: (10.1038/sj.jid.5700850) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Rab11 is associated with corneosesmosin-positive lamellar granules. Double staining of Rab11 (5-nm gold labels, small arrows) and corneodesmosin (10-nm gold labels, large arrows) using cryo-ultra microtomy. (a) A lower magnification view of the stratum corneum (C), the most superficial stratum granulosum (G1), the second (G2), the third (G3), and the fourth stratum granulosum (G4) is shown. The rectangular area marked in (a) is shown at higher magnification in (b) Nu, nucleus. (a) Bar=2μm. (b) Bar=100nm. Journal of Investigative Dermatology 2007 127, 2166-2170DOI: (10.1038/sj.jid.5700850) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 Rab11 is associated with cathepsin D-positive lamellar granules. Double staining of Rab11 (5-nm gold labels, small arrows) and cathepsin D (10-nm gold labels, large arrows) using a cryo-ultra microtomy method. (a) A lower magnification view of the most superficial stratum granulosum (G1), the second (G2), and the third (G3) stratum granulosum is shown. The rectangular area marked in (a) is shown at higher magnification in (b). (a) Bar=1μm. (b) Bar=100nm. Journal of Investigative Dermatology 2007 127, 2166-2170DOI: (10.1038/sj.jid.5700850) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 6 Rab11 is associated with glucosylceramide-positive lamellar granules. Double staining of Rab11 (5-nm gold labels, small arrows) and glucosylceramides (10-nm gold labels, large arrows) using cryo-ultra microtomy. The rectangular area marked in (a) is shown at higher magnification in (b). d, desmosome. (a) Bar=500nm. (b) Bar=100nm. Journal of Investigative Dermatology 2007 127, 2166-2170DOI: (10.1038/sj.jid.5700850) Copyright © 2007 The Society for Investigative Dermatology, Inc Terms and Conditions