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Apoptosis Assay

KK Kenji Kawada
TS Takehiko Sasazuki
SS Senji Shirasawa
SH Suguru Hasegawa
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In terminal deoxynucleotidyl transferase-mediated dUTP-FIFC nick end-labeling (TUNEL) assay, cells were cultured under glutamine-free DMEM (25 mM glucose, 0 mM glutamine) containing 10% FBS for 72 hours and then fixed in 4% parafolmaldehyde. The number of TUNEL-positive cells in each of the three slides was counted in five random fields (magnification, ×400) by software BZ-H3C (Keyence) and expressed as a percentage of the total cells counted. The activity of caspase-3 and -7 was measured by using caspase-Glo 3/7 assay (Promega) according to the manufacturer's protocol. Caspase activity was normalized to the cell number counted by CCK-8 cell proliferation assay under the same density and conditions.

Copyright and License information:  ©2016 The Authors
This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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