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. 2019 Nov 15;10:5176. doi: 10.1038/s41467-019-13150-0

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© The Author(s) 2019

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. 7 — Progenitors with active Tlr2 locus labeled at E8.5 contribute to adult hematopoiesis. a Tlr2^CreERT2Rosa26^EYFP embryos pulsed with a single dose of 4-OHT between E6.75-E10.5 were monitored postpartum for the presence of Tlr2^CreERT2EYFP⁺ cells in their peripheral blood (PB) up to 16 weeks of age (mean ± SEM; n = 9–25; * indicates short-term hematopoietic potential which persisted for only the first 4 weeks of life. b Percentage of Tlr2^CreERT2EYFP⁺ cells, labeled at E8.5, and their contribution to the main CD45⁺ lineages in different hematopoietic organs is shown (mean ± SEM; n = 5–7; see gating strategy in Supplementary Fig. 8a). c Tlr2^CreERT2Rosa26^EYFP embryos were pulsed with a single dose of 4-OHT at E8.5, and labeling efficiency of indicated hematopoietic populations was analyzed in E15.5 FL by FCM. LT-HSC gate is shown as an example. d Tlr2^CreERT2Rosa26^EYFP embryos were pulsed with a single dose of 4-OHT at E8.5. At 16 weeks of age, Lin-depleted BM (CD45.2) (w16 Lin^– BM; see Supplementary Fig. 8b) was transferred to lethally irradiated primary recipients (CD45.1/2) along with support BM cells (0.5 × 10⁶ CD45.1), and the level of Tlr2^CreERT2EYFP⁺ cells was monitored in PB of primary recipients until week 16 (upper left panel). The percentage of Tlr2^CreERT2EYFP⁺ cells, their organ distribution and fate in primary recipients is shown (mean ± SEM; n = 6). Bar graphs show the reconstitution success and fate of Tlr2^CreERT2EYFP⁺ cells in primary recipients (A–C’) of BM from eight individual donors (#1–8). Sorted Tlr2^CreERT2EYFP⁺ cells (see Supplementary Fig. 8c) from Lin-depleted BM of 16-week old primary recipients were transferred to secondary, lethally irradiated recipients (CD45.1/2) along with support BM cells (0.5 × 10⁶, CD45.1) and the frequency of Tlr2^CreERT2EYFP⁺ cells in PB was monitored until week 16 (the outcome of BMT from one primary recipient (donor T) is shown (secondary recipients, upper row, left panel). The percentage of Tlr2^CreERT2EYFP⁺ cells, their organ distribution and fate in secondary recipients is shown (mean ± SEM; n = 8). Bar graphs on the bottom indicate the reconstitution success and fate of Tlr2^CreERT2EYFP⁺ cells in secondary recipients (1´−34´) of BM from four primary recipients. Source data are provided as a Source Data file