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. 2018 Jan 9;8:199. doi: 10.1038/s41598-017-18606-1

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Expression of marine luciferases does not alter target cells sensitivity to cytotoxic agents. (A) LDH release assay. K562 cells stably expressing Gluc, Nluc, Tluc16, Mluc7, and parental line were plated in a 96-well U bottom plate in phenol red free media, followed by treatment with media alone (control), digitonin (30 µg/ml for 90 minutes), or 1% Triton X-100 (TX100) for 45 minutes. Post incubation the plates were spun at 250 g for 4 minutes. Supernatants were carefully transferred into a flat bottom plate followed by the calorimetric detection of LDH activity as described in the manufacturer protocol. (B) Calcein release assay. Indicated cell lines were pre-incubated with Calcein-AM as described in methods section and the experiment set up was same as for LDH release assay, except that the supernatants were transferred into a black walled flat bottom plate followed by measurement of fluorescence (Excitation 485 ± 9 nm, Emission 530 ± 9 nm) in a Biotek Synergy plate reader.