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. 2024 May 30;25(6):1083–1096. doi: 10.1038/s41590-024-01844-7

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© The Author(s) 2024

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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Extended Data Fig. 8 — a, Schematic of adoptive transfer of CD45.2⁺ 10E8-UCA^H B cells into CD45.1⁺ wild-type mice followed by immunization with 10E8-GT10.2 12mer with alhydrogel and analysis until 21 DPI. b, Quantification of percent of B cells in GC of spleen from mice immunized with 10E8-GT10.2 12mer at 7, 14, and 21 DPI, as in a. Green lines show the mean percentage of 10E8-UCA^H GC B cells in mice immunized with 10E8-GT10.2 12mer and gray lines show mean percentage of cells immunized with 10E8-GT9-KO 12mer (n = 3). Data from one representative of three independent experiments shown. c, Quantification of epitope specific CD45.2⁺10E8-UCA^H in mice immunized with 10E8-GT10.2 12mer at different time points, as in a. Error bars indicate mean ± SD from mice in each group (n = 3). Data from one representative of three independent experiments shown. d, ELISA of Ab response of 10E8-UCA^H to immunizations with 10E8-GT10.2 12mer, as in a. Green dots represent IgG titer specific to 10E8-GT10.2 probe and gray dots represent IgG level to KO probe. Data pooled from three independent experiments. Each symbol represents a different mouse. Bars indicate geometric mean and geometric SD from mice in all three groups. n = 4-5 mice in each group. e, Graph showing the MFI of mAb-GT10.2-CD45.2 10E8-UCA^H-14DPI-PE with varying amounts of competing antibodies (µg) on x-axis (yellow: CD45.2 Ab; red: CD45.1 Ab). Dotted line represents the ‘no competition’ control. f, Representative flow cytometric plots of CD45.2⁺10E8-UCA^H B cells recruitment to GCs 7 and 14 DPI in spleens of mice transferred to reach frequencies of ~1:10⁴ 5:10⁴ and 25:10⁴ CD45.2⁺10E8-UCA^H precursors and immunized with 5 μg of 10E8-GT10.2 12mer with alhydrogel or 10E8-GT9-KO 12mer with alhydrogel (control). g, Quantification of CD45.2⁺ in GC in spleen of mice immunized in f. Data are pooled from two independent experiments. Error bars indicate mean ± SD from mice in each group (n = 3 mice/independent group).