Evidence of Heterogeneity and Quantitative Differences of the Type 1 5α-Reductase Expression in Cultured Human Skin Cells – Evidence of its Presence in.

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Evidence of Heterogeneity and Quantitative Differences of the Type 1 5α-Reductase Expression in Cultured Human Skin Cells – Evidence of its Presence in Melanocytes Wenchieh Chen, Christos C. Zouboulis, Martin Fritsch, Ulrike Blume-Peytavi, Vitam Kodelja, Sergij Goerdt, Van Luu-The, Constantin E. Orfanos Journal of Investigative Dermatology Volume 110, Issue 1, Pages 84-89 (January 1998) DOI: 10.1046/j.1523-1747.1998.00080.x Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 Results of immunocytochemistry using rabbit anti-serum raised against human type 1 5α-reductase. (a) Adult facial sebocytes; (b) neonatal foreskin keratinocytes; (c) adult nongenital keratinocytes (flank); (d) occipital dermal papilla cells; (e) beard dermal papilla cells; (f) occipital fibroblasts; (g) melanocytes; (h) dermal microvascular endothelial cells; (i) chin fibroblasts. Scale bars,50 mm. Journal of Investigative Dermatology 1998 110, 84-89DOI: (10.1046/j.1523-1747.1998.00080.x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Immunocytochemistry of adult facial sebocytes from the fourth passage in culture. The smaller, less differentiated sebocytes are more strongly stained with anti-type 1 5α-reductase antibody as compared with larger, well-differentiated ones. Scale bar, 50 mm. Journal of Investigative Dermatology 1998 110, 84-89DOI: (10.1046/j.1523-1747.1998.00080.x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Results of western blot study with 12.5% sodium dodecyl sulfate/polyacrylamide gel electrophoresis. With the exception of beard dermal papilla cells, all the cells under examination show two closely lying bands in the range of molecular weight 21–27kDa. Journal of Investigative Dermatology 1998 110, 84-89DOI: (10.1046/j.1523-1747.1998.00080.x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 4 Results of northern blot study with 1.2% MOPS/fomaldehyde agarose gel. Quantitative differences are observed among different cell types, after correction by β-actin and G3PDH, respectively, which show a parallel relationship. Journal of Investigative Dermatology 1998 110, 84-89DOI: (10.1046/j.1523-1747.1998.00080.x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 5 (a) DNA standards; (b) β-actin and type 1 5α-reluctase PCR products. (a) HPLC analysis of PCR products from 22 cycles amplification. cDNA of β-actin and type 1 5α-reductase were amplified in the same tube, whereby the concentration of β-actin primers was one-fourth of that of type 1 5α-reductase primers (0.125 μM vs 0.5 μM). (b) Elution of PCR products was correlated to size in accordance with HPLC analysis of DNA standards. Journal of Investigative Dermatology 1998 110, 84-89DOI: (10.1046/j.1523-1747.1998.00080.x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 6 Linear correlations between PCR cycles and area under concentration of HPLC analysis. The appropriate cycles for type 1 5α-reductase and β-actin were shown to be 22–28 and 14–20, respectively. The value of type 1 5α-reductase in neonatal foreskin keratinocytes, when corrected with β-actin, is about 1.5–10 higher that of adult facial sebocytes. Journal of Investigative Dermatology 1998 110, 84-89DOI: (10.1046/j.1523-1747.1998.00080.x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions