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. 1999 Nov 29;147(5):1049–1062. doi: 10.1083/jcb.147.5.1049

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© 1999 The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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Embryonic motoneurons express Fas and FasL both in situ and after purification. Total RNA was prepared from the indicated rat tissues, and RT-PCR analysis followed by Southern blotting was performed using specific primers and internal probes for FasL (A) and Fas (B). RT-PCR for β-actin was used as an internal control to ensure that similar concentrations of mRNA were present in all samples. For each mRNA, the first panel corresponds to positive and negative control samples prepared from indicated tissues of adult rat; the results are in agreement with several reports in the literature. The second panel shows results from E14 rat embryos, using either freshly dissected ventral spinal cord or motoneurons purified by a metrizamide-immunoaffinity method in conditions that limit de novo synthesis of mRNA. Fas and FasL are present at significant levels in motoneurons. Alternate unlabeled lanes in all panels are control in which reverse transcriptase was omitted from the incubation.