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. 2000 Jun 12;149(6):1215–1224. doi: 10.1083/jcb.149.6.1215

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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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graphic file with name JCB0002104.f6a.jpg — Exposure of PE on the surface of R-41 mutant cells. a, Binding of ¹²⁵I-SA-Ro to the plasma membrane. CHO-K1 and R-41 mutant cells were incubated in ethanolamine-deficient medium without (CHO-K1, R-41) or with either 30 μM PE (R-41+PE) or 20 μM ethanolamine (R-41+Etn) at 39.5°C for 3 d. Cells were incubated with ¹²⁵I-SA-Ro (30,000 cpm/ml) on ice for 30 min. The cells were then washed and their radioactivity was measured. The results show the mean ± SEM (n = 3). b, Exposure of PE on the cleavage furrow membrane of R-41 mutant cells. R-41 mutant cells were incubated in ethanolamine-deficient medium with (R-41+Etn) or without (R-41) 20 μM ethanolamine at 39.5°C for 3 d and then synchronized in mitotic phase. The synchronized cells were collected, washed, and then further incubated for 30 min. The cells were washed and stained with FL–SA-Ro for 30 min on ice, then washed and photographed. Bar, 10 μm. The arrowheads indicate the cleavage furrow.

graphic file with name JCB0002104.f6b.jpg — Exposure of PE on the surface of R-41 mutant cells. a, Binding of ¹²⁵I-SA-Ro to the plasma membrane. CHO-K1 and R-41 mutant cells were incubated in ethanolamine-deficient medium without (CHO-K1, R-41) or with either 30 μM PE (R-41+PE) or 20 μM ethanolamine (R-41+Etn) at 39.5°C for 3 d. Cells were incubated with ¹²⁵I-SA-Ro (30,000 cpm/ml) on ice for 30 min. The cells were then washed and their radioactivity was measured. The results show the mean ± SEM (n = 3). b, Exposure of PE on the cleavage furrow membrane of R-41 mutant cells. R-41 mutant cells were incubated in ethanolamine-deficient medium with (R-41+Etn) or without (R-41) 20 μM ethanolamine at 39.5°C for 3 d and then synchronized in mitotic phase. The synchronized cells were collected, washed, and then further incubated for 30 min. The cells were washed and stained with FL–SA-Ro for 30 min on ice, then washed and photographed. Bar, 10 μm. The arrowheads indicate the cleavage furrow.