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. 2020 Apr 28;11:2054. doi: 10.1038/s41467-020-15937-y

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© The Author(s) 2020

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. 8 — a Histograms showing upregulation of activation markers HLA-DR, CD86 and CD83 in in vitro-differentiated cDC2 in response to TLR4 (LPS) and TLR8 (VTX-2337) overnight (16 h) stimulation. b Representative FACS plots and quantification of mixed lymphocyte reaction (MLR) using in vitro and in vivo differentiated cDC2, pDC, and pre/AS-DC. FACS-sorted DC subsets were activated overnight (16 h) using a TLR agonist cocktail (LPS 10 ng/ml, R848 1 μg/ml and Poly(I:C) 25 μg/ml) and co-cultured with CTV-labeled naive T cells for 7 days (n = 2 independent donors for pDC generated in vivo and pre/AS-DC generated in vitro and in vivo and n = 3 independent donors for in vitro and in vivo generated cDC2 and in vitro generated pDC in two independent experiments). **p < 0.01, one-way ANOVA test with Holm–Sidak’s multiple comparisons. c Intracellular flow cytometry analysis of TNFα and IL-12 expression in in vitro and in vivo-differentiated DC subsets upon overnight (16 h) stimulation with TLR agonist cocktail (LPS 10 ng/ml, R848 1 μg/ml, and Poly(I:C) 25μg/ml) as compared to unstimulated cells (NT). n = 4 independent cord blood donors. **p < 0.01, ***p < 0.001, one-way ANOVA test Holm–Sidak’s multiple comparisons. Data are presented as floating bars ranging from min to max and line represents median b or as bar graphs with mean ± SEM (c).