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. 2000 Mar 20;191(6):977–984. doi: 10.1084/jem.191.6.977

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© 2000 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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Binding of Stat5 to the bcl-x promoter in Mo7e-Neo and Mo7e-p210 cell lines. Cells were treated for 3 h with CGP 57148 or tyrphostin AG 555, and formation of Stat5-DNA complexes was determined by an EMSA using a radiolabeled bcl-x probe. DNA binding activity of Spi-1/PU.1 in Mo7e-p210 cells was also analyzed as a specificity control. Nuclear extracts from untreated cells (C) were preincubated with antibodies specific for Stat5 (+anti-ST5) or Spi-1 (anti-Spi1), and with an excess of an unlabeled bcl-x probe as a specific competitor (+SP) or with an irrelevant nonspecific probe (+non-SP). Extracts from COS cells transfected with a Spi-1 expression vector (COS-Spi) were used as positive controls.