Detection of c-sis proto-oncogene transcripts by direct enzyme-labeled cDNA probes and in situ hybridization

J. T. McClintock, I. J. Chan, S. R. Thaker, A. Katial, F. E. Taub, A. E. Aotaki-Keen, Leonard M Hjelmeland

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6 Scopus citations

Abstract

Using in situ hybridization and platelet-derived growth factor (PDGF) cDNA probes labeled with horseradish peroxidase, PDGF-A and -B (c-cis proto- oncogene) mRNA transcripts were identified and localized in proliferating cultures. A human retinal pigment epithelial (RPE) cell line and a glial cell line were treated with either transforming growth factor beta-1 (TGFB1), phorbol-12-myristate-13-acetate (PMA), or thrombin from human plasma and compared for their ability to stimulate the production of PDGF-A and -B. Expression of both PDGF-A and -B transcripts were found to be localized predominantly in the cytoplasm of TGFB1-treated RPE cells, with a portion of these cells displaying a hybridization response in the nuclear region. When compared to PMA- and thrombin-treated cells, TGFB1 stimulated the RPE cell line to yield the greatest amount of detectable PDGF mRNA. In addition, the hybridization response observed in TGFB1-treated cells was shown to be RNA dependent.

Original languageEnglish (US)
Pages (from-to)102-108
Number of pages7
JournalIn Vitro Cellular and Developmental Biology - Animal
Volume28 A
Issue number2
StatePublished - 1992
Externally publishedYes

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ASJC Scopus subject areas

  • Cell Biology
  • Clinical Biochemistry
  • Developmental Biology

Cite this

McClintock, J. T., Chan, I. J., Thaker, S. R., Katial, A., Taub, F. E., Aotaki-Keen, A. E., & Hjelmeland, L. M. (1992). Detection of c-sis proto-oncogene transcripts by direct enzyme-labeled cDNA probes and in situ hybridization. In Vitro Cellular and Developmental Biology - Animal, 28 A(2), 102-108.