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. 2022 Mar 24;12:5130. doi: 10.1038/s41598-022-09051-w

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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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Subtle difference in the composition of the most toxic 6mer seed between human and mouse cancer cell lines. (A) Results of the 4096 6mer seed duplex screen in a human (H4) and a mouse (GL261) glioma cell line. Cells were reverse transfected in triplicates in 384 well plates with 10 nM of individual siRNAs. The cell viability of each 6mer seed duplex was determined by quantifying cellular ATP content 96 h after transfection. All 4096 6mer seeds are ranked by the average effect on cell viability of the two cell lines from the most toxic (left) to the least toxic (right). We consider an siRNA highly toxic if it reduces cell viability 90% or more and moderately toxic if it reduces cell viability 50% or more (black stippled line). (B) Average seed composition of the 100 most (left) and 100 least (right) toxic 6mer seeds in H4 and GL261 cells screened with 4096 6mer seed containing siRNAs. (C) Left: Regression analysis showing correlation between the 6mer Seed Tox observed in H4 cells and the average of the two previously tested human cell lines HeyA8 and H460. Right: Regression analysis showing correlation between the 6mer Seed Tox observed in GL261 cells and the average of the two previously tested murine cell lines 3LL and M565. p-values were calculated using Pearson correlation analysis. (D) Seed composition of the 100 most toxic 6mer seeds in three human (top) and three murine (bottom) cell lines. (E) Average most toxic seed composition of all human (top) and all murine (bottom) cell lines. p-value is derived from a multinomial mixed effects regression model.