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Hung-Yi Su, Jonathan G. H. Hickford, Pauline H. B. The, Anne M

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1 Increased Vibrissa Growth in Transgenic Mice Expressing Insulin-like Growth Factor 1 
Hung-Yi Su, Jonathan G.H. Hickford, Pauline H.B. The, Anne M. Hill, Roy Bickerstaffe  Journal of Investigative Dermatology  Volume 112, Issue 2, Pages (February 1999) DOI: /j x Copyright © 1999 The Society for Investigative Dermatology, Inc Terms and Conditions

2 Figure 1 Analysis of transgenic status by PCR in IGF-1 transgenic mice. Genomic DNA was extracted from mouse tails and amplified by PCR as outlined in Materials and Methods. Lane 1, a molecular weight size marker, lambda DNA digested with HindIII;lanes 2–13, DNA from 12 transgenic mice of line 1417 after PCR amplification and four (lanes 5, 6, 8, and 11) showed 350 bp UHS-KER-IGF-1 fragments;lane 14, DNA from one nontransgenic mouse of line 1377;lane 15, DNA from one nontransgenic mouse of line 1455;lane 16, DNA from one transgenic mouse of line 1455 and the 350 bp fragment is shown. Journal of Investigative Dermatology  , DOI: ( /j x) Copyright © 1999 The Society for Investigative Dermatology, Inc Terms and Conditions

3 Figure 2 Analysis of IGF-1 mRNA expression in the skin of mice from line 1417 by RPA. RPA was accomplished by hybridization of total RNA extracted from mouse skin with a radioactively labeled anti-sense IGF-1 riboprobe. The ovine IGF-1 probe protected a 287 bp fragment (lanes 6–8 and 10) corresponding to mRNA transcribed from the transgene (IGF-1 and vector) as described in Materials and Methods. Lane 1, partially degraded IGF-1 probe, which may be due to radiolytic decay;lane 2, a 330 nt mouse actin probe was prepared by in vitro transcription of a 250 bp mouse β-actin DNA (provided with the RPA kit, Ambion) by T7 RNA polymerase according to the methods described in SP6/T7 Transcription Kit, Boehringer (Mannheim, Germany);lane 3, a 250 bp fragment corresponding actin message was protected by actin probe and detected in mouse liver;lane 4, some background is shown on the hybridization of IGF-1 probe with yeast RNA and this probably due to incomplete digestion of the probe by RNase;lane 5, blank lane;lanes 6–9, IGF-1 probe hybridized with transgenic mouse skin RNA from line 1417;lane 10, IGF-1 probe hybridized with mouse skin RNA from one expressor of line 1453. Journal of Investigative Dermatology  , DOI: ( /j x) Copyright © 1999 The Society for Investigative Dermatology, Inc Terms and Conditions

4 Figure 3 Measurement of vibrissa growth at days 11–16 and 16–21 of age in one of the transgenic line 1417 mice using 35S-cystine incorporation into hair fibers. Three spots (from top) represent incorporation of 35S-cystine into fibers of mouse vibrissae at days 11, 16, and 21 of age. Journal of Investigative Dermatology  , DOI: ( /j x) Copyright © 1999 The Society for Investigative Dermatology, Inc Terms and Conditions

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