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. 2022 Sep 16;20:144. doi: 10.1186/s12964-022-00901-8

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© The Author(s) 2022

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 6 — TRAF6 is essential for the Notch1-mediated inflammatory response and ROS generation in LPS-stimulated macrophages. BMMs were isolated from the Notch1^FL/FL and Notch1^M−KO mice and transfected with CRISPR/Cas9-mediated TRAF6 activation or TRAF6 KO vector. A Western blot analysis of TRAF6, p-TAK1 and p-P65 in BMMs transfected with CRISPR/Cas9-mediated TRAF6 activation and control. Representative of three experiments. B Quantitative RT-PCR analysis of TNF-α, IL-1β, CCL-2 and CXCL-10 mRNA levels in vitro (n = 3–4 samples/group). C Detection of ROS production by Carboxy-H2DFFDA in LPS-stimulated macrophages from the Notch1^FL/FL mice. Quantification of ROS-producing Macrophages (green) (n = 4–6 mice/group). Scale bars, 40 μm. D Western blot analysis of TRAF6, p-TAK1 and p-P65 in BMMs transfected with CRISPR/Cas9-mediated TRAF6 KO and control. Representative of three experiments. E Quantitative RT-PCR analysis of TNF-α, IL-1β, CCL-2 and CXCL-10 mRNA levels in vitro (n = 3–4 samples/group). F Detection of ROS production by Carboxy-H2DFFDA in LPS-stimulated macrophages from the Notch1^M−KO mice. Quantification of ROS-producing Macrophages (green) (n = 4–6 mice/group). Scale bars, 40 μm. All data represent the mean ± SD. *p < 0.05. **p < 0.01. ***p < 0.001