Identification of cDNA Encoding a Serine Protease Homologous to Human Complement C1r Precursor from Grafted Mouse Skin Sung June Byun, Young Yil Bahk,

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Identification of cDNA Encoding a Serine Protease Homologous to Human Complement C1r Precursor from Grafted Mouse Skin Sung June Byun, Young Yil Bahk, Zae Young Ryoo, Kyoung-Eun Kim, Ha-Young Hwang, Jung-Woong Lee, Jong-Yuk Yi, Tae-Yoon Kim Journal of Investigative Dermatology Volume 116, Issue 3, Pages 374-379 (March 2001) DOI: 10.1046/j.1523-1747.2001.01257.x Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Northern blot analysis of the clone homologous to human complement Clr precursor. (A) Total RNAs were prepared from normal, autografted, and allografted mouse skin biopsies at days 1 and 8 post-transplantation. Representative hybridization signals were obtained from mC1r at the expected position corresponding to an mRNA size of 3 kb. The hybridization signal for GAPDH is shown as a control at the same blot. (B) Values for mC1r mRNA levels in relation to GAPDH mRNA levels in the three groups of animals. Lane 1, normal mouse skin; lane 2, autograft day 1; lane 3, autograft day 8; lane 4, allograft day 1; lane 5, allograft day 8. Journal of Investigative Dermatology 2001 116, 374-379DOI: (10.1046/j.1523-1747.2001.01257.x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Nucleotide and deduced amino acid sequences of the cDNA homologous to human complement Clr precursor. The cDNA was screened from the mouse skin cDNA library. The putative signal peptide and polyadenylation signal sequences are indicated by underlining. The three boxed Asn residues are the possible glycosylation sites. The sequence is available from the GenBank, accession number AF148216 Journal of Investigative Dermatology 2001 116, 374-379DOI: (10.1046/j.1523-1747.2001.01257.x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Sequence alignment of the amino acid sequences of the isolated mouse cDNA and human complement Clr precursor. Amino acids that are physicochemically similar are indicated by a plus symbol. Dashes in the sequences represent gaps introduced to maximize alignment. Amino acid residues marked with closed diamonds represent key amino acids conserved in trypsin-type serine protease. The solid arrow indicates the putative Arg-Ile peptide bond cleaved during the activation of mC1r. Journal of Investigative Dermatology 2001 116, 374-379DOI: (10.1046/j.1523-1747.2001.01257.x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Expression of mC1r in normal skin tissue and allografted skin tissue on day 8 post-transplantation. The skin biopsies harvested from normal (a, b) and allografted (c, d) mice were evaluated by immunohistochemistry using an anti-mC1r polyclonal antibody, which was raised against the recombinant mC1r protein, and preimmune rabbit serum as a control antibody [e (normal skin tissue) and f (allograft skin tissue 8 d post-transplantation)]. Magnification: a, c, 40× b, d, e, f, 200×. Journal of Investigative Dermatology 2001 116, 374-379DOI: (10.1046/j.1523-1747.2001.01257.x) Copyright © 2001 The Society for Investigative Dermatology, Inc Terms and Conditions