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Fig. 1. Generation of Hr^-/- mice. (A) Expression of Hr mRNA in Hr mutant mice. Northern analysis of mRNA from backskin of adult (10 week old) mice. hr, Hr^hr allele (HRS/J); hr^rh, Hr^rh-j allele (RHJ/LeJ); +, wild type allele. Asterisks indicate high molecular weight RNA species detected only in Hr^hr allele. (B) Expression of HR protein in Hr mutant mice. Western analysis of extracts from adult backskin shows protein detected by HR-specific antibody (-Hr). hr, Hr allele (SKH2/J); hr^rh, Hr^rh allele (CBA-hr^rh); +, wild type allele; Ponceau, Ponceau Red stained membrane. Molecular weight is shown in kDa. (C) Targeting vector used to create Hr null (Hr^-) allele. Exons 8-10 were replaced with a neo cassette (PGKneo) using homologous recombination. Exons are not drawn to scale. RI, EcoRI sites. (D) Southern analysis of mice with targeted gene deletion. Southern blot of EcoRI-digested genomic DNA. Probe is region indicated by black bar in C. Band of 9 kb represents wild-type allele; 6 kb band represents recombinant allele; +, wild type allele; -, recombinant (Hr^-) allele. (E) Hr mRNA is disrupted in mice carrying targeted gene deletion. Northern analysis of RNA prepared from skin of P15 mice. +/+, wild type; -/-, Hr^-/-. (F) Mice carrying targeted gene deletion do not express HR protein. Western analysis of extracts from skin with HR-specific antibody (-Hr). (G) Skin wrinkling in Hr^-/- mouse (1 year old). (H) Comparison of 1-year-old Hr mouse mutants. Null, Hr^-/- mouse; rhino, Hr^rh/Hr^rh mouse (CBA-Hr^rh); Hr, Hr^hr/Hr^hr mouse (SKH2/J).

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