HKAP1.6 and hKAP1.7, Two Novel Human High Sulfur Keratin-Associated Proteins are Expressed in the Hair Follicle Cortex Yutaka Shimomura, Noriaki Aoki,

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hKAP1.6 and hKAP1.7, Two Novel Human High Sulfur Keratin-Associated Proteins are Expressed in the Hair Follicle Cortex Yutaka Shimomura, Noriaki Aoki, Masaaki Ito Journal of Investigative Dermatology Volume 118, Issue 2, Pages 226-231 (February 2002) DOI: 10.1046/j.0022-202x.2001.01653.x Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 hKAP1.6 gene and its mRNA can be detected in the human genome and in hair follicle mRNA. (A) PCR amplification of the hKAP1.1 A/B and hKAP1.6 genes in three individuals (lanes 1–3). (B) RT-PCR amplification of hKAP1.1 A/B and hKAP1.6 mRNAs from human hair follicles (lane 1). GAPDH mRNA, 597 bp in size, was amplified as a control using two primers located, respectively, in exon 3 and exon 7 of GAPDH gene. A genomic fragment of GAPDH, 1101 bp in size, was not detected (lane 3). PCR without reverse transcription did not detect any products (lanes 2, 4), indicating that there was no genomic DNA contamination in this experiment. RT(+)/RT(–) denote controls with or without reverse transcription. MMW, molecular weight marker. Journal of Investigative Dermatology 2002 118, 226-231DOI: (10.1046/j.0022-202x.2001.01653.x) Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Nucleotide sequence of hKAP1.6 and hKAP1.7, and the deduced amino acid sequence of the protein products. (A) hKAP1.6; (B) hKAP1.7. Nucleotides are numbered consecutively at the left-hand side. The derived amino acid sequence appears below the nucleotide sequence and the pentapeptide repeats are boxed. Polyadenylation signal is underlined. The accession numbers for the respective nucleotide sequences are: hKAP1.6, AB052868; hKAP1.7, AB055057. Journal of Investigative Dermatology 2002 118, 226-231DOI: (10.1046/j.0022-202x.2001.01653.x) Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 The predicted amino acid sequences of hKAP1.6 and hKAP1.7 and multialignment of KAP1 family members. Residues that are identical are colored light blue. Dashes denote gaps in the sequence to maximize alignment. The pentapeptide repeats are boxed using blue lines. Brackets show the five KAP subdomains. Note that the N-terminal domain of hKAP1.6 is boxed with a red line. The accession numbers for the respective human, rat, and sheep protein sequences are: hKAP1.1 A (hB2A), X63337; hKAP1.1B (AC007455, nt 8801–9334); hKAP1.2 (hB2B), X63338; sB2A, kr2a_sheep.sw; sB2B, kr2b_sheep.sw; sB2C, kr2c_sheep.sw: sB2D, kr2d_sheep.sw; rB2E and rB2F, AB003753. Journal of Investigative Dermatology 2002 118, 226-231DOI: (10.1046/j.0022-202x.2001.01653.x) Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 A probe common to hKAP1.1 A/B, hKAP1.6, and hKAP1.7 hybridizes to mRNA transcripts present in the cortex of the hair follicle. Due to the high DNA sequence homology of these genes, the in situ probe used in this experiment recognizes all four transcripts. (A) Longitudinal hair follicle section; (B) higher magnification of (A); (C) longitudinal section of a hair follicle possessing a medulla. The red arrows demarcate the area of major KAP expression. Note the lack of inner root sheath (B) and medulla (C) labeling. ors, outer root sheath; irs, inner root sheath; co, cortex; dp, dermal papilla; med, medulla, hcu, cuticle of the hair shaft; icu, cuticle of the inner root sheath. Scale bar: 150 µm. Journal of Investigative Dermatology 2002 118, 226-231DOI: (10.1046/j.0022-202x.2001.01653.x) Copyright © 2002 The Society for Investigative Dermatology, Inc Terms and Conditions