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. 2022 Feb 9;13:762. doi: 10.1038/s41467-022-28454-x

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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 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. 4 — A Dual luciferase reporter assay showing luciferase activities of THP-1 cells transfected with random short Lp-RNA (ctrlRNA), RsmY, or tRNA-Phe. Results are shown as relative luciferase activity (ratio Firefly/Renilla activity), where luciferase activity of ctrlRNA is defined as 100%. Bars indicate the (mean) SD of eight independent experiments. For statistical analysis, a two-way ANOVA was performed with p < 0.0001 (****). Source data provided as Source data file. B Dual luciferase reporter assay showing luciferase activities of THP-1 cells in absence (NI) or presence of wt or a ∆rsmY Lp-EVs. Lp-EVs affect protein expression depending on the presence of the un-translated regions (UTR) of ddx58 (RIG-I), irak1, or rel (cRel). Results are shown as relative luciferase activity (ratio Firefly/Renilla activity), luciferase activity of THP-1 cells (NI) was defined as 100%. Bars show the (mean) SD of n = 7 independent experiments. Statistical analysis, a two-way ANOVA with p < 0.05 significant (*), p < 0.01 very significant (**) and p < 0.001 extremely significant (***) was performed. Source data provided as Source data file. C Relative luciferase activities of THP-1 cells transfected with RsmY or tRNA-Phe. RsmY and tRNA-Phe affect protein expression significantly less in presence of an Ago2-Inhibitor. The bars indicate the (mean) SD of three independent experiments. Statistical analysis, a two-way ANOVA analysis was performed with p < 0.0001 (****). Source data provided as Source data file. D Dual luciferase reporter assay showing the relative luciferase activities (ratio Firefly/Renilla activity) of THP-1 cells incubated with Lp-EVs+/− Ago2-Inhibitor. When inhibiting Ago2, the suppressing effect of Lp-EVs on relative luciferase activity depending on the presence of the un-translated regions (UTR) of ddx58 (RIG-I), irak1 or rel (cRel) is significantly reduced. The bars show the (mean) SD of n = 4 independent experiments. Statistical analysis, a two-way ANOVA with values of p < 0.05 significant (*), p < 0.01 very significant (**) and p < 0.001 extremely significant (***) and p < 0.0001 (****) was performed. Source data provided as Source data file.