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Figure 5. Yeast cells expressing HsCen3p present a large bud. A, Yeast strain expressing HsCen3p or Cdc31p were grown in inducing condition (raffinose + galactose), repressing condition (glucose), or in raffinose medium (the Gal promotor is neither repressed or induced). The number of cells were determined at hourly intervals. Cells expressing HsCen3p stopped growing in raffinose + galactose and in raffinose medium at 6 and 10 h, respectively. B, Proteins from yeast strain expressing HsCen3p growing in glucose, raffinose, or raffinose + galactose medium were prepared and analyzed by Western blot with anti-HsCen3p antibodies. HsCen3p is rapidly and efficiently induced in raffinose + galactose and a very low signal is detected in raffinose after a 12-h culture. C, Yeast strain expressing HsCen3p was grown in inducing condition (raffinose + galactose). The number of unbudded, small-budded, or large-budded cells were determined at hourly intervals. Cells presenting a large bud start accumulating at 6 h. D, Phase-contrast microscopy of yeast strain expressing HsCen3p grown in inducing conditions. Most of the cells present a large bud. Bar, 10 μm. E, FACS analysis of yeast strain expressing HsCen3p or Cdc31p grown in inducing conditions (raffinose + galactose). At 12 h, cells expressing HsCen3p have mainly a G2 DNA content, whereas control cells have a G1 DNA content. Image published in: Middendorp S et al. (2000) © 2000 The Rockefeller University Press. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license Larger Image Printer Friendly View

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