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Multiparameter analysis of apoptosis in puromycin-treated Saccharomyces cerevisiae

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Abstract

In Saccharomyces cerevisiae, a typical apoptotic phenotype is induced by some stress factors such as sugars, acetic acid, hydrogen peroxide, aspirin and age. Nevertheless, no data have been reported for apoptosis induced by puromycin, a damaging agent known to induce apoptosis in mammalian cells. We treated S. cerevisiae with puromycin to induce apoptosis and evaluated the percentage of dead cells by using Hoechst 33342 staining, transmission electron microscopy (TEM) and Annexin V flow cytometry (FC) analysis. Hoechst 33342 fluorescence images were processed to acquire parameters to use for multiparameter analysis [and perform a principal component analysis, (PCA)]. Cell viability was evaluated by Rhodamine 123 (Rh 123) and Acridine Orange microscope fluorescence staining. The results show puromycin-induced apoptosis in S. cerevisiae, and the PCA analysis indicated that the increasing percentage of apoptotic cells delineated a well-defined graph profile. The results were supported by TEM and FC. This study gives new insights into yeast apoptosis using puromycin as inducer agent, and PCA analysis may complement molecular analysis facilitating further studies to its detection.

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Conflict of interest

The authors declare no conflicts of interest.

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Authors and Affiliations

  1. Department of Biomolecular Sciences, University of Urbino “Carlo Bo”, Urbino, Italy

    Barbara Citterio, Maria Cristina Albertini, Marco B. L. Rocchi & Elena Piatti

  2. Department of Biology, University of Rome “Tor Vergata”, Rome, Italy

    Lina Ghibelli

  3. Department of Earth, Life and Environment Sciences, University of Urbino “Carlo Bo”, Urbino, Italy

    Elisabetta Falcieri, Michela Battistelli & Barbara Canonico

  4. UTAPRAD-DIM, ENEA Frascati, Rome, Italy

    Laura Teodori

  5. Department of Life and Environmental Sciences, Polytechnic University of Marche, Ancona, Italy

    Maurizio Ciani

Authors
  1. Barbara Citterio
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  2. Maria Cristina Albertini
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  3. Lina Ghibelli
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  4. Elisabetta Falcieri
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  5. Michela Battistelli
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  6. Barbara Canonico
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  7. Marco B. L. Rocchi
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  10. Elena Piatti
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Correspondence to Elena Piatti.

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Communicated by Olaf Kniemeyer.

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Citterio, B., Albertini, M.C., Ghibelli, L. et al. Multiparameter analysis of apoptosis in puromycin-treated Saccharomyces cerevisiae . Arch Microbiol 197, 773–780 (2015). https://doi.org/10.1007/s00203-015-1110-7

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  • DOI: https://doi.org/10.1007/s00203-015-1110-7

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