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. 2022 May 25;13:2921. doi: 10.1038/s41467-022-30658-0

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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. 7 — a Evolution of OC43 S genes. Neighbor-Joining phylogeny constructed from an alignment of S genes of North-American/Eurasian field variants with bovine coronavirus as outgroup (see Supplementary Table 2). Branches were re-ordered from top to bottom based on collection date with the year of isolation indicated by color-coding in a sidebar (right) to visualize S evolution among and within distinct S lineages. Branchpoint confidence values >80 are given for all major nodes. The topological positions of S types selected for comparative analysis of antigenic properties are indicated by red dots; that of the oldest OC43 isolate, the USA/1967 strain, is indicated by a green star. Strain and GenBank identifiers of selected S types: NL/2019 (OK245434); BE/2004 (AY9034551); USA/1990(A-II) (KF530088); NL/2004 (OK245433); USA/2016 (MF314143.1); USA/1967 (AAT84354.1); USA/1990(B) (KF530065.1). b Spike protein sequence variability and position of neutralizing epitopes. One protomer in the OC43 spike trimer structure (PDB: 6OHW) is colored according to amino acid conservation across 228 OC43 S sequences. Ribbon diagrams of 46C12, 43E6 and 37F1 are overlaid and shown as a silhouette. In the right panel, two of the protomers are transparent and the shielded epitope region for 56E10, 45B9, and 65A11 are circled. GenBank identifiers of OC43 spike protein sequences are provided in Supplementary Fig. 18b. c, d Heatmap showing ELISA binding signal (OD450_nm) of S1-directed mAbs (H2L2 format, 0.4 μg/ml) to (mutant) S1 proteins of OC43 isolates (c) or S1 proteins of multiple betacoronaviruses including human OC43, bovine BCoV, porcine PHEV and equine ECoV (all belonging to the Betacoronavirus-1 species within the Embecovirus subgenus), as well as the more distantly related embecoviruses MHV and HKU1, the merbecovirus MERS-CoV and two sarbecoviruses SARS-CoV and SARS-CoV-2 (d), with S1 of the orthologous USA/1967 as a reference. The antibody specificity (S1 domain and antibody competition group) is indicated on the left and neutralizing antibodies are highlighted in orange, similar as in Fig. 1c. Data from a representative experiment are shown.