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. 2017 Nov 13;8:1457. doi: 10.1038/s41467-017-01388-5

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© The Author(s) 2017

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. 1 — PLCγ1/PLCγ2 double deficiency blocks B cell development at the pre-pro-B cell stage. BM from Plcg1 ^+/+ Plcg2 ^+/+, Plcg1 ^−/− Plcg2 ^+/+, Plcg1 ^+/+ Plcg2 ^−/−, or Plcg1 ^−/− Plcg2 ^−/− mice were transplanted into lethally irradiated congenic wild-type CD45.1⁺ mice. Six to eight weeks after transplantation, the recipients were analyzed. a BM cells from the recipients are largely YFP⁺ donor-derived. Lin⁻CD45.2⁺ BM cells from the recipients were analyzed. Numbers indicate percentages of YFP⁺ cells in the gated live cells. b Both PLCγ1 and PLCγ2 are deleted in YFP⁺ BM cells from Plcg1 ^−/− Plcg2 ^−/− mice. YFP⁺ BM cells from the recipients were subjected to western blot analysis. c Both PLCγ1 and PLCγ2 are expressed in pro-B cells. Sorted pro- (B220^loIgM⁻CD43⁺) and FO (B220⁺IgM^loCD23⁺CD21^int) B cells from wild-type mice were subjected to western blot analysis. d BM cells from the recipients were stained with anti-CD45.2, anti-B220, and anti-IgM. Numbers indicate percentages of pro/pre-, immature, and mature B cells in the gated CD45.2⁺ population. e Bar graphs show the numbers of CD45.2⁺ pro/pre-, immature, and mature B cells in BM. f BM cells from the recipients were stained with anti-CD45.2, anti-B220, anti-IgM, anti-CD25, and anti-CD43. Numbers indicate percentages of pro-B (upper) and pre-B (lower) cells in the gated CD45.2⁺B220⁺IgM⁻ population. g Bar graphs show the numbers of CD45.2⁺ pro-B cells and pre-B cells in BM. h BM cells from the recipients were stained with anti-CD45.2, anti-B220, anti-CD43, anti-CD24, and anti-BP-1. BM from μMT and Jak3-deficient mice served as the controls. Numbers indicate percentages of pre-pro-B (fraction A), early pro-B (fraction B) and late pro-B/early pre-B (fraction C/C′) cells in the gated CD45.2⁺B220⁺CD43⁺ pro-B cell population. i Bar graphs show the numbers of CD45.2⁺ fractions A, B, and C/C′ B cells in BM. Error bars show ± SEM. Data shown are obtained from or representative of 10 (a, d–g) or 8 (h, i) mice of each genotype or representative of 2 (b, c) independent experiments. Control data are representative of 3 μMT and Jak3-deficient mice