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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. 1 — a Flow-cytometry profiling of extra- and intravascular CD19⁺ cells in WT C57BL/6 mouse kidney, urinary bladder, liver, lung, spleen, peritoneal lavage and blood (kidneys n = 9, other organs n = 7). Data representative of three independent experiments. b Absolute extravascular B cell counts per gram of tissue across organs shown in a. ^xPeritoneal B cell counts calculated per single lavage and adjusted for total volume of PBS injected. c Percentage of B cells within live extravascular CD45⁺ cell subset across organs shown in a. Kidney ^*P = 0.0156, bladder ^*P = 0.0156, liver ^*P = 0.0312, lung ^*P = 0.0156. d Proportion of naïve (IgD⁺IgM⁺), IgM⁺IgD^- and switched (double negative, DN) B cells from intra- (left) and extravascular (right) compartments in organs shown in a, n = 7 per group, means with SEM shown. The difference in naïve B cells proportions between blood and a tested organ is compared; intravascular: kidney P = 0.2188, bladder ^*P = 0.0156, liver P = 0.1562, lung P = 0.3750, peritoneal P = 0.6250, spleen P = 0.0781; extravascular: ^*P = 0.0156. e Flow-cytometry identification of CD45⁺CD3^-CD19⁺ cells in matched kidney and spleen from deceased human organ donors (left) and their phenotyping based on expression of IgD and CD27 (middle). Percentage of naïve (IgD⁺CD27^-), switched memory (IgD^-CD27⁺), DN and double positive cells within CD45⁺CD3^-CD19⁺ cell subset shown (right) (n = 10 per group, means with SEM shown). The difference in naïve B cells proportions between kidney and spleen is tested, ^*P = 0.0137. f Tukey plots show percentage of B-1a (CD19⁺CD23^-CD5⁺) and CD9⁺ cells in extravascular IgD^- B cell compartments of organs described in a. Data pooled from three independent experiments at two independent SPF animal facilities (total n = 12 per organ); %B-1: kidney ^***P = 0.0010, bladder, liver and lung ^***P = 0.0005; %CD9⁺ cells: kidney ^**P = 0.0024, bladder ^***P = 0.0005, liver P = 0.4785, lung ^**P = 0.0029. g Confocal microscopy of adult (left) and neonatal (right) renal cortex from WT C57BL/6 mice focusing on extravascular localization of CD5⁺ B cells. Sections stained for CD19 (red), CD5 (green) and CD31 (grey). Representative of three independent experiments. Blue arrows point at extravascular CD19⁺CD5⁺ B cells. Blood samples represent always only intravascular cells. P values were calculated using two-tailed Wilcoxon matched-pairs signed rank test (c-f). Bar plots show medians unless stated otherwise. Source data are provided with this paper.