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. Author manuscript; available in PMC: 2009 Jun 8.

Published in final edited form as: Exp Cell Res. 2008 Jan 16;314(6):1237–1249. doi: 10.1016/j.yexcr.2007.12.028

Fig. 2 — Ca²⁺ induces DNA cleavage and histone release in isolated nuclei from non-apoptotic thymocytes. (a) Induction of DNA cleavage in nuclei with 0.5 mM CaCl₂ treatment. 1× 10⁴ nuclei were treated with a fluorometric TUNEL assay (right) and counter stained with DAPI (left). The data shown represents one of three independent experiments. (b) Induction of DNA degradation with various concentrations of CaCl₂ ranging from 0.1 μM to 1 mM. C is a control. Nuclei (2 × 10⁶) were incubated in vitro at 37°C for 90 min with or without CaCl₂ as described in Materials and Methods. Nuclei were harvested and treated with protease K and RNase. DNA (2 μg) from the nuclei was run on an agarose gel and stained with ethidium bromide. Typical internucleosomal DNA ladders were observed starting around 50 μM CaCl₂. The data shown represents one of at lease three independent experiments. (c) Divalent cations and Ca²⁺ specific DNA degradation in isolated nuclei. EDTA and EGTA block Ca²⁺ dependent DNA degradation in nuclei obtained from thymocytes. The data shown represents one of two independent experiments.

Fig. 2 — Ca²⁺ induces DNA cleavage and histone release in isolated nuclei from non-apoptotic thymocytes. (a) Induction of DNA cleavage in nuclei with 0.5 mM CaCl₂ treatment. 1× 10⁴ nuclei were treated with a fluorometric TUNEL assay (right) and counter stained with DAPI (left). The data shown represents one of three independent experiments. (b) Induction of DNA degradation with various concentrations of CaCl₂ ranging from 0.1 μM to 1 mM. C is a control. Nuclei (2 × 10⁶) were incubated in vitro at 37°C for 90 min with or without CaCl₂ as described in Materials and Methods. Nuclei were harvested and treated with protease K and RNase. DNA (2 μg) from the nuclei was run on an agarose gel and stained with ethidium bromide. Typical internucleosomal DNA ladders were observed starting around 50 μM CaCl₂. The data shown represents one of at lease three independent experiments. (c) Divalent cations and Ca²⁺ specific DNA degradation in isolated nuclei. EDTA and EGTA block Ca²⁺ dependent DNA degradation in nuclei obtained from thymocytes. The data shown represents one of two independent experiments.