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. 2023 Jul 11;28(12):5380–5389. doi: 10.1038/s41380-023-02166-0

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© The Author(s) 2023

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. 5 — Conditioned media were collected from PBMCs after treatment with 1 μM TLR9 agonist ODN2216 (Donor#_TLR9; containing mixture of TLR9-signaling induced cytokines) or vehicle (Donor#_CT) and were diluted 10-fold to pre-treat human iPSC-derived microglial cultures for ~14 h. After the pre-treatment, microglial cultures were subjected to LPS priming for 4 h and nigericin stimulation (inflammasome activation) for 2 h. The supernatants from microglial cultures were collected for cytokine measurement by MSD multiplex assay. The concentrations of measured cytokines are plotted in chart (A) IL-1β, (B) TNFα, (C) IL-10, and the CellTiter measurements are plotted in chart (D). PBMCs were prepared from four healthy donors (Donor 1-Donor 4). Biological replicates from n = 3 independent experiments (each experiment with 3 biological replicates) are plotted in small-sized orange squares, mean values of biological replicates per experiment are plotted in black dots. Mean ± SD, One-way ANOVA with Fisher’s LDS pairwise comparisons of mean values from independent experiments. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ns (not significant).