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. 2015 Sep 28;1:15037. doi: 10.1038/cddiscovery.2015.37

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Copyright © 2015 Cell Death Differentiation Association

This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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DIDS binds irreversibly to HeLa cells and make them fluorescent. (a) DIDS fluorescence signal as a function of concentration shows a biphasic behavior. DIDS concentrations higher than 50 μM displayed a reduction in fluorescence signal most likely due to internal quenching. (b) HeLa cells preincubated with DIDS (5, 50 or 500 μM) for 2 h were washed and fluorescence due to DIDS (420 nm) was determined (n=3). HeLa cells autofluorescence was substracted from the signal obtained for those preincubated with DIDS. Collectively, these data suggest that preincubating cells with 50 μM DIDS leads to an accumulation of this inhibitor of at least 5 μM inside cells.