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Spheroids were grown for 5 days as indicated; at T4, cells were collected and cytocentrifuged at 800 rpm for 5 min on glass coverslip and next fixed in 3.7% paraformaldehyde 20 min. Nuclei were stained with 2.25 µg/mL of Hoechst 33258. Cells with apoptotic nuclear bodies were counted in 12 different fields on 6 different glass coverslips/condition by using a 20X objective. Results were expressed as percentage of apoptotic cells over the field.
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Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
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