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. 2020 Mar 10;11:1297. doi: 10.1038/s41467-020-15061-x

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© The Author(s) 2020

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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Fig. 5 — a Schematic diagrams of apoptosis-regulating circuits. Apoptosis is induced by only the translational activation of Bax (left) or both the translational activation of Bax and translational repression of Bcl-xL (right). b The viability of cells transfected with apoptosis-regulating circuits. HeLa cells were co-transfected with 1xMS2(U)site2-Bax (cap analog: A-cap), 2xScMS2(C)-BclxL (cap analog: ARCA), and CaVT or MS2CP mRNAs. All mRNAs contained N1mΨ. Cell viability was analyzed by the WST-1 assay. The bar graph shows the means ± SD. (n = 4 independent wells in each transfection condition). ***P < 0.001 compared to no Bax mRNA/no CaVT or MS2CP samples by ANOVA with Dunnett’s multiple comparison test (two-sided). Exact P values are shown in Supplementary Table 1. Source data are provided as a Source Data file. c, d Annexin V (apoptosis marker) and SYTOX Red (dead cell marker) staining. HeLa cells were co-transfected with 1xMS2(U)site2-Bax mRNA (cap analog: A-cap), 2xScMS2(C)-BclxL mRNA (cap analog: ARCA), and CaVT mRNA. For the positive control, 1xMS2(U)site2-Bax mRNA (cap analog: ARCA) was transfected. All mRNAs contained N1mΨ. One day after the transfection, the cells were stained and analyzed by a flow cytometer. The bar graph shows the average of four independent experiments (mean ± SD) (c). Representative two-dimensional dot plots (d). **P < 0.01, ***P < 0.001 compared to the negative control by ANOVA with Dunnett’s multiple comparison test (two-sided). Exact P values are shown in Supplementary Table 1. Source data are provided as a Source Data file.