Characterization of the Human Hair Shaft Cuticle–Specific Keratin-Associated Protein 10 Family Hiroki Fujikawa, Atsushi Fujimoto, Muhammad Farooq, Masaaki.

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Characterization of the Human Hair Shaft Cuticle–Specific Keratin-Associated Protein 10 Family Hiroki Fujikawa, Atsushi Fujimoto, Muhammad Farooq, Masaaki Ito, Yutaka Shimomura Journal of Investigative Dermatology Volume 133, Issue 12, Pages 2780-2782 (December 2013) DOI: 10.1038/jid.2013.233 Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 The expression of hair keratin-associated protein (KRTAP) 10 in the human hair shaft and cultured cells. (a) Western blot with an anti-KRTAP10 antibody. The antibody recognized KRTAP10 proteins from human hairs. (b) Indirect immunofluorescence (IIF) of human hair shafts with the anti-KRTAP10 antibody showed the expression of KRTAP10 proteins in the upper keratinizing zone of the hair shaft cuticle. (c) Double IIF studies with the anti-KRTAP10 and anti-keratin 85 (K85) antibodies showed that the expression of KRTAP10 and K85 proteins finely overlapped in the cuticle layer. (d–f) IIF studies in HaCaT cells in which both hemagglutinin (HA)-tagged KRTAP10-1 (HA-KRTAP10-1) and Flag-tagged K82 were overexpressed. (g–i) IIF studies in HaCaT cells in which only HA-KRTAP10-1 was overexpressed. Counterstaining with 4′,6-diamidino-2-phenylindole is shown in blue (b, c, f, and i). Scale bars=100μm (b) and 10μm (d). Journal of Investigative Dermatology 2013 133, 2780-2782DOI: (10.1038/jid.2013.233) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Characterization of human KRTAP10-1 protein. Co-immunoprecipitation (co-IP) between hemagglutinin (HA)-KRTAP10-1 and Flag-KRTAP10-1. (b) Co-IP between HA-KRTAP10-1 and K85. (c) Co-IP between HA-KRTAP10-1 and Flag-K32, and between HA-KRTAP10-1 and Flag-tagged K82 (Flag-K82). The cell lysate was precipitated with anti-HA agarose beads (a–c). (d) Co-IP between HA-KRTAP10-1 and Flag-K10, and between HA-KRTAP10-1 and Flag-K32. (e) Co-IP between HA-KRTAP10-1 and Flag-tagged truncated K82 proteins. Full-length K82 (Flag-K82-F) was used as a positive control. Amino acid residues for each domain of K82 are shown on the top panel. The cell lysate was precipitated with anti-Flag agarose beads (d, e). (f) GST pull-down assays. GST-Flag fusion proteins for truncated K82 and HA-KRTAP10-1 proteins were overexpressed in competent cells and HEK293T cells, respectively. Schematic representation of each fusion protein is shown on the top panel. WB, western blot. Journal of Investigative Dermatology 2013 133, 2780-2782DOI: (10.1038/jid.2013.233) Copyright © 2013 The Society for Investigative Dermatology, Inc Terms and Conditions