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. 2023 Nov 4;14:7081. doi: 10.1038/s41467-023-42625-4

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© The Author(s) 2023

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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Fig. 2 — a Schematic showing parabiosis experimental setup (left): Congenically marked WT animals were connected for 5 weeks before organs harvesting. Flow cytometry (middle) and quantification (right) of peripheral blood CD19⁺ lymphocytes chimerism in each parabiont, (n = 6 per group). Data representative of two independent experiments. b Percentages of host-derived extravascular IgM⁺IgD^-, DN, naïve and B-1a B cells found in kidney, liver, lung, female reproductive tract (FRT), spleen and blood from parabiosis experiment described in a. P values were calculated with two-tailed Wilcoxon matched-pairs signed rank test: ^*P = 0.0312, FRT & Fat (B-1a) P = 0.0625, spleen (B-1a) P = 0.1250, blood (IgM⁺IgD^-) P = 0.8438, blood (B-1a) P = 0.9999. c Schematic showing B-cell depletion experimental setup (left): WT C57BL/6 mice were injected with anti-mouse CD20 depleting antibody or isotype control. On day 7, EdU was introduced into their drinking water for 21 days, after which organs (incl. blood) were harvested. Percentage of B cells in peripheral blood leukocytes from these mice was checked both on day 7 and 28 (right, n = 5 per group). Data representative of two independent experiments. d Absolute extravascular CD19⁺ cell counts found in kidney (controls n = 10, depleted n = 9, ^****P < 0.0001), urinary bladder (n = 5, P = 0.5476), liver (n = 5, ^**P = 0.0079), lung (n = 5, P = 0.6905), peritoneal wash (n = 5, ^**P = 0.0079) and spleen (n = 5, ^**P = 0.0079) from depleted mice or controls on day 28 as described in c. P values were calculated with two-tailed Mann-Whitney U test. e Percentage of EdU positive cells in extravascular naïve, IgM⁺IgD^-, DN and B^-1a B cell compartment found in organs from circulating B-cell depleted mouse cohort as described in c. P values were calculated with paired two-tailed t test (data normality confirmed with Shapiro-Wilk test): Kidney (n = 8) ^***P = 0.0003 (IgM⁺IgD^-), ^***P = 0.0002 (DN), ^***P = 0.0004 (B-1a); bladder (n = 5) ^***P = 0.0005 (IgM⁺IgD^-), ^*P = 0.0313 (DN), ^***P = 0.0010 (B-1a); liver (n = 5) ^*P = 0.0193 (IgM⁺IgD^-), ^*P = 0.0119 (B-1a); lung (n = 5) ^***P = 0.0006 (IgM⁺IgD^-), ^***P = 0.0004 (B-1a); peritoneal (n = 5) ^***P = 0.0001 (IgM⁺IgD^-), ^**P = 0.0060 (DN), ^***P = 0.0006 (B-1a); spleen (n = 5) ^**P = 0.0047 (IgM⁺IgD^-), ^**P = 0.0089 (DN), ^**P = 0.0055 (B-1a). Representative flow cytometry plots are shown in Supplementary Fig. 2d. All box plots show medians. Source data are provided with this paper.