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. 2022 Nov 5;12:18803. doi: 10.1038/s41598-022-23650-7

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© The Author(s) 2022

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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BCL-2 family members and P53 protein expression levels. Expression levels of (a, c) BCL-2 family members, including BCL-X_L (anti-apoptotic), BCL-2 (anti-apoptotic), MCL-1 (anti-apoptotic), BAX (pro-apoptotic), NOXA (pro-apoptotic) and PUMA (pro-apoptotic) (a) or P53 (c) were analyzed by western blot in H9 and FN2.1 cells at 4, 8 and 24 h upon 1% O₂ hypoxia treatment. ACTIN was used as loading control. Representative blots of three independent experiments are shown (original western blot images are presented in Supplementary Fig. S13, S14, and S15). Bar graphs represent the densitometric quantification of bands. Data are expressed as means + SEM fold induction relative to normoxia (Norm.) and statistical analysis was done by Student’s t-test, (*) p < 0.05, (**) p < 0.01, and (***) p < 0.001 versus normoxia. (b) mRNA expression levels of MCL-1 were analyzed by RT-qPCR at 2, 4, 8, and 24 h upon 1% O₂ hypoxia induction. RPL7 expression was used as normalizer. Graph shows mean + SEM mRNA fold induction relative to normoxia (arbitrarily set as 1) from three independent experiments. Statistical analysis was done by Student’s t-test.