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. 2021 Jul 27;12:4559. doi: 10.1038/s41467-021-24876-1

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© The Author(s) 2021

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 — Concentrations of human pro-inflammatory factors in (a) blood serum of mice at euthanasia or (b) in cultured media 120 h post-transduction. For (a), each dot represents a mouse: untreated (UT, black, n = 2), GFP (green, n = 13), wtBRAF (blue, n = 4), BRAF^V600E (red, n = 13). For (b), wtBRAF (blue), and BRAF^V600E (red), four donors for each group. Data are presented as box plots minimum to maximum with lines representing means. c Correlation between blood cytokine levels and survival of BRAF^V600E mice (n = 11). Non-linear regression one-phase decay. The goodness of fit: R². d Growth curve of HSPCs exposed to CM from wtBRAF or BRAF^V600E HSPCs. n = 9 for three donors/group. Data are presented as mean values +/− SD. e Clonogenic potential of HSPCs exposed to CM from wtBRAF or BRAF^V600E HSPCs. n = 7 for three donors/group. Data are presented as mean values +/− SEM. f–h Growth curves of HSPCs exposed to cytokines: (f) IL1α (light green: 0.5 ng/ml, medium green: 5 ng/ml and dark green: 50 ng/ml), (g) IL1β (light blue: 0.5 ng/ml, medium blue: 5 ng/ml and dark blue: 50 ng/ml) and (h) TNFα (light red: 0.5 ng/ml, medium red: 5 ng/ml and dark red: 50 ng/ml). Technical replicates: CTRL (n = 5), IL1α-IL1β-TNFα (n = 4). CTRL values (black) are repeated in f, g, and h. Data are presented as mean values +/− SD. Statistical test: linear mixed-effects model (LME), followed by post hoc analysis at 24 and 192 h. See also Source Data file. i Clonogenic potential of HSPCs treated with recombinant cytokines for 96 h. CTRL(n = 4), IL1α (n = 4), IL1β (n = 4), TNFα (n = 4). Data are presented as mean values +/− SEM. j Gene expression analysis of p21, IL1α, MCP1, and IL8 upon cytokine treatments (three donors for: CTRL, IL1α0.5, IL1α5, IL1α50, IL1β0.5, and TNFα0.5; four donors for: IL1β5, IL1β50, TNFα5, and TNFα50). Data are presented as mean +/− SD. k Representative images of a skin lesion from an histiocytosis patient stained for TNFα. Tissue morphology analyzed by hematoxylin and eosin (H/E). Scale bars: 100 or 200 µm. Statistical tests: for (b), two-tailed Mann−Whitney; for a, i, j, Kruskal−Wallis with post-Dunn’s multiple comparisons. ns non significant; *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.