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. 2021 Sep 15;11:18345. doi: 10.1038/s41598-021-97869-1

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© The Author(s) 2021

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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

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CSE treatment and SCAL1 expression enables A549 cells to resist apoptotic induction with MSB. (a) Annexin V-Alexa Fluor 488-gated events representing apoptotic cells were unaffected by CSE treatment, except for cells treated with 1× CSE. (b) Live cell populations (AV−/PI−) remained unchanged across different CSE treatments, except for the 1× CSE setup exhibiting a shift to more apoptotic (AV+) cells. (c) Low caspase 3/7 levels persisted in different CSE treatments but increased in the 1× CSE setup. (d) Mitochondrial integrity measured with TMRM intensity was unaffected by incubation with lower levels CSE but was greatly reduced in cells treated with 1× CSE. (e) Annexin V-Alexa Fluor 488-gated events were reduced upon SCAL1 overexpression. (f) Live cell population (AV−/PI−) increase was accompanied by a decrease of apoptotic cell (AV+) number upon SCAL1 upregulation. (g) Annexin V-Alexa Fluor 488-gated events increased upon SCAL1 knockdown. (h) SCAL1 knockdown resulted in a reduced live cell population (AV−/PI−) and a greater number of apoptotic (AV+) cells. (i,j) Fluorescent staining of caspase 3/7 and active mitochondria showed lower caspase 3/7 levels and a higher TMRM intensity in SCAL1-overexpressing cells. (k,l) SCAL1 knockdown resulted in higher caspase 3/7 staining intensity and lower TMRM levels. Data presented are representative of three independent trials performed in triplicates and are expressed as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001.