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. 2022 Apr 27;605(7909):372–379. doi: 10.1038/s41586-022-04677-2

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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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Extended Data Fig. 2 — (a) Nucleosides of S. tokodaii tRNA^Val2/3 were subjected to LC/MS analysis. Top panel shows the UV trace at 254 nm. Second to bottom panels show XICs detecting proton adducts of modified nucleosides as indicated. N³²⁴ nucleoside was not detected in the bottom panel. m¹G is derived from tRNA^Val2, which was co-isolated with tRNA^Val3 (Supplementary Fig. 1). Asterisks indicate unassigned ions. (b) RNA-MS of RNase T₁ digests of the isolated S. tokodaii tRNA^Val2/3. The upper panel shows the BPC of RNase T₁-digested fragments. The sequence and molecular mass of each RNA fragment numbered in BPC are listed in Supplementary Table 1a. The lower panel shows the XIC of the divalent negative ion of the fragment containing N³²⁴. (c) RNA-MS of RNase A digests of isolated S. tokodaii tRNA^Val2/3. The upper panel shows the BPC of RNase A-digested fragments. The sequence and molecular mass of each RNA fragment numbered in BPC are listed in Supplementary Table 1b. The lower panel shows the XIC of the divalent negative ion of the fragment containing N³²⁴.