Peroxiredoxin is Ubiquitously Expressed in Rat Skin: Isotype-Specific Expression in the Epidermis and Hair Follicle Jeong Eun Lee, Byung Dae Kwon, Jee-Bum.

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Peroxiredoxin is Ubiquitously Expressed in Rat Skin: Isotype-Specific Expression in the Epidermis and Hair Follicle Jeong Eun Lee, Byung Dae Kwon, Jee-Bum Lee, Young-Ho Won, Young Pio Kim Journal of Investigative Dermatology Volume 115, Issue 6, Pages 1108-1114 (December 2000) DOI: 10.1046/j.1523-1747.2000.00177.x Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Classification of the Prx family. Journal of Investigative Dermatology 2000 115, 1108-1114DOI: (10.1046/j.1523-1747.2000.00177.x) Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Prx antibodies show monospecificity. Twenty micrograms of authentic Prx isotypes I, II, or III were resolved on 12% sodium dodecyl sulfate–polyacrylamide gels and separately reacted with the three isotype-specific antibodies. Journal of Investigative Dermatology 2000 115, 1108-1114DOI: (10.1046/j.1523-1747.2000.00177.x) Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Three Prx isotypes are ubiquitously expressed in rat skin. Twenty micrograms of rat skin extracts from whole skin, epidermis, and the dermis were subjected to 12% sodium dodecyl sulfate–polyacrylamide gel electrophoresis and western blotting followed by incubation with polyclonal antibodies against isotype-specific anti-Prx antibodies. The purified authentic markers for Prx isotypes were also loaded with samples. Lane A, authentic marker; lane B, whole skin extract; lane C, dermal extract; lane D, epidermal extract. Identical results were obtained in three experiments. Journal of Investigative Dermatology 2000 115, 1108-1114DOI: (10.1046/j.1523-1747.2000.00177.x) Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Prx I–III isotypes are ubiquitously expressed in rat keratinocytes and fibroblasts. Twenty micrograms of cell extracts from rat keratinocytes and fibroblasts were subjected to 12% sodium dodecyl sulfate–polyacrylamide gel electrophoresis and western blotting followed by incubation with polyclonal antibodies against isotype-specific anti-Prx antibodies. The purified authentic markers for Prx isotypes were also loaded with samples. Lane A, authentic marker; lane B, cell extract from rat keratinocytes; lane C, cell extract from rat fibroblasts. Identical results were obtained in three experiments. Journal of Investigative Dermatology 2000 115, 1108-1114DOI: (10.1046/j.1523-1747.2000.00177.x) Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 Prx is mainly localized to the cytoplasm of rat keratinocytes and fibroblasts. Rat keratinocytes (a) and fibroblasts (b) were fixed in methanol and acetone, and immunofluorescence labeling was performed with anti-Prx II antibody. Confocal laser images of rat keratinocytes (a) and fibroblasts (b) are shown. Scale bar: 10 μm. Journal of Investigative Dermatology 2000 115, 1108-1114DOI: (10.1046/j.1523-1747.2000.00177.x) Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 6 Immunohistochemical staining for Prx expression in rat skin. Formalin-fixed rat skin was stained with isotype-specific anti-Prx antibodies as described in Materials and Methods. Prx expression in the epidermis (a–c) and hair follicle (d–f): Prx I (a, d); Prx II (b, e); and Prx III (c, f). Scale bar: 50 μm. Journal of Investigative Dermatology 2000 115, 1108-1114DOI: (10.1046/j.1523-1747.2000.00177.x) Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 7 Immunofluorescence staining for TR and Trx expression in rat skin. Frozen sections of rat skin were stained with isotype-specific anti-TR and Trx antibodies as described in Materials and Methods. TR expression in the epidermis (a) and hair follicle (b), and Trx expression in the epidermis (c) and hair follicle (d). Scale bar: 50 μm. Journal of Investigative Dermatology 2000 115, 1108-1114DOI: (10.1046/j.1523-1747.2000.00177.x) Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 8 In situ hybridization of Prx II mRNA in rat skin. Rat skin fixed in paraformaldehyde was hybridized with the anti-sense probe for Prx II cRNA. The positive signals were shown in the whole skin (a), epidermis (b), and hair follicles (c). Rat skin was also hybridized with the sense probe for Prx II (d). Black dots delineate dermoepidermal junction and junction between the dermis and hair follicles. Scale bar: 50 μm. Journal of Investigative Dermatology 2000 115, 1108-1114DOI: (10.1046/j.1523-1747.2000.00177.x) Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 9 Time-course change of Prx II expression in UVB-irradiated rat skin. Prx II expression levels were examined in nonirradiated normal skin (lane C) and UVB-irradiated skin from 0 to 24 h postirradiation (lanes 0–24 h). M indicates an authentic marker for Prx II. Journal of Investigative Dermatology 2000 115, 1108-1114DOI: (10.1046/j.1523-1747.2000.00177.x) Copyright © 2000 The Society for Investigative Dermatology, Inc Terms and Conditions