Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2024 Feb 9;15:1229. doi: 10.1038/s41467-024-45201-6

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 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/.

PMC Copyright notice

Fig. 8 — a Flow cytometry plots of IgM vs IgD staining of splenic B cells representative of C57BL/6 (n = 7), Tlr3^−/−Tlr7^−/−Tlr9^−/− (n = 7), EGT-315 B6 (n = 6), EGT-315 Tlr3^−/−Tlr7^−/−Tlr9^−/− (n = 6). Squares: mature B cells (left), transitional B cells (right upper), immature B cells (right lower). Summary of 9 experiments. b Follicular (gate upper middle) and marginal zone (gate lower right) B cell distribution in Tlr deficient mice. Exemplary flowcytometry of spleen cells from C57BL/6 (n = 3), Tlr3^−/−Tlr7^−/−Tlr9^−/− (n = 2), EGT-315 B6 (n = 6), EGT-315 Tlr3^−/−Tlr7^−/−Tlr9^−/− (n = 5) using the B cell markers CD23 and CD21. Summary of 3 experiments. c GFP-specific IgM measured by GFP specific ELISA from individual mice: C57BL/6 (mean = 1160, SD = 476, n = 4), Tlr3^−/−Tlr7^−/−Tlr9^−/− (mean = 392, SD = 229, n = 6), EGT-315 B6 (mean = 3249, SD = 2037, n = 29), EGT-315 Tlr3^−/−Tlr7^−/−Tlr9^−/− (mean = 312, SD = 171, n = 8). Red lines depict mean value. EGT-315 B6 vs. EGT-315 Tlr3^−/−Tlr7^−/−Tlr9^−/− ***P = 0.0004. Ordinary one-way ANOVA Tukey’s multiple comparisons test. d Survival/proliferation of purified B cells isolated from individual mice 72 h after in vitro stimulation (no stimulus, LPS = Tlr4 ligand, anti-IgM = BCR ligand, R848 = Tlr7 ligand, 1668 PTO = Tlr9 ligand) measured by flow cytometry using propidium iodide to distinguish live vs dead cells. C57BL/6 (black bar, for anti-IgM: mean = 15.1, SD = 5.9, n = 9), Tlr3^−/−Tlr7^−/−Tlr9^−/− (red bar, for anti-IgM: mean = 3.2, SD = 2.0, n = 9), EGT-315 B6 (green bar, for anti-IgM: mean = 18.6, SD = 6.3, n = 8), EGT-315 Tlr3^−/−Tlr7^−/−Tlr9^−/− (blue bar, for anti-IgM: mean = 4.0, SD = 3.0, n = 7). Summary of 12 experiments, 2 way ANOVA, Sidak´s multiple comparisons test ****P < 0.0001. e Role of Btk in survival/proliferation of purified B cells 72 h after in vitro stimulation. Anti-IgM was used with and without the addition of the Btk-inhibitor Ibrutinib. Cells from individual mice were tested. C57BL/6 and EGT-315 B6 combined (green bar, for anti-IgM: mean = 11.9, SD = 4.3, n = 6), Tlr3^−/−Tlr7^−/−Tlr9^−/− and EGT-315 Tlr3^−/−Tlr7^−/−Tlr9^−/− B6 combined (blue bar, for anti-IgM: mean = 1.7, SD = 1.1, n = 6). Summary of 5 experiments, 2-way ANOVA, Sidak´s multiple comparisons test, ****P < 0.0001. f Tlr3^−/−Tlr7^−/−Tlr9^−/− B cells induced by LPS (Tlr4-ligand) upregulate CD23. Upper panels, LPS-mediated in vitro induction of CD23 is higher in Tlr3^−/−Tlr7^−/−Tlr9^−/− B cells compared to wild type B cells. Flowcytometry examples of stimulated B cells from C57BL/6 and Tlr3^−/−Tlr7^−/−Tlr9^−/− mice for 72 h with medium alone (red), LPS (10 μg/ml, blue) and LPS + IL-4 (LPS 10 μg/ml+ IL-4 250 U/ml, green). Right panels, Summary of MFI of CD23 from three independent experiments. Mann-Whitney test Two-tailed pairwise comparison ***P = 0.0007 and **P = 0.0031. B6 (n = 10), Tlr3^−/−Tlr7^−/−Tlr9^−/− (n = 7). Source data are provided as a Source Data file.