Multiparametric staining to identify apoptotic human cells

Claudia Negri, Maddalena Donzelli, Rosa Bernardi, Laura Rossi, Alexander Bürkle, A. Ivana Scovassi

Research output: Contribution to journalArticlepeer-review

Abstract

To analyze relevant features of HeLa and HL60 cells driven into apoptosis by etoposide, we have developed a new 'tricolor' assay, based on the simultaneous analysis in the single cell of chromatin condensation, DNA degradation, and cellular poly(ADP-ribose) synthesis. The latter reaction is catalyzed by poly(ADP-ribose)-polymerase (E.C. 2.4.2.30), an enzyme which is activated by the presence of DNA free ends. The protocol consists in the visualization of apoptotic cells by Hoechst staining, TUNEL assay, and immunoreaction with anti-poly(ADP-ribose) antibody. We thus provide the first evidence that endogenous poly(ADP-ribose) production is indeed stimulated in cells undergoing apoptosis after treatment with antitumoral drugs, and that the monitoring of this endogenous enzymatic reaction, combined with morphological and other biochemical parameters, should facilitate the detection of apoptotic cells.

Original languageEnglish
Pages (from-to)174-177
Number of pages4
JournalExperimental Cell Research
Volume234
Issue number1
DOIs
Publication statusPublished - Jul 10 1997

ASJC Scopus subject areas

  • Cell Biology

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