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. 2022 Sep 13;13:5347. doi: 10.1038/s41467-022-32928-3

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© The Author(s) 2022

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 — a Volcano plot showing significantly upregulated and downregulated (p-adj < 0.05 and log₂fc > 0.5) genes in IL-1β (5 ng/mL) treated MSCs (n = 2) compared to PBS treated MSCs (n = 2). b Gene-set enrichment analyses (GSEA) show significant increase in expression of genes related to cytokine production, inflammatory response, apoptosis and regulation of cell-cell adhesion in IL-1β treated MSCs (n = 2) compared to PBS treated MSCs (n = 2). Enrichment plots with normalized enrichment score (NES) and false discovery rate (FDR) are shown. c Heat map of selected genes upregulated in IL-1β treated MSCs (n = 2) compared to PBS treated MSCs (n = 2) with expression changes more than 1.5-fold (FDR < 0.05). d RT-qPCR validation of increased mRNA expression of Il1b (n = 4, 4 biological replicates), Il1a (n = 4, 4), Il6 (n = 3, 3), Ccl2 (n = 3, 3), Ccl5 (n = 3, 3), Tlr2 (n = 4, 4) and Myd88 (n = 3, 3) in IL-1β treated MSCs compared with PBS treated MSCs. Data were normalized with Hprt1 expression. Data are shown in bar graphs as mean ± SEM. Statistical significances were determined using two-tailed unpaired t-test. e, f BM MSCs were treated with vehicle (PBS), IL-1α (5 ng/mL) or IL-1α (5 ng/mL) + IL-1R1 Ab (1 μg/ml) (n = 4 biological replicates per group) (e) and vehicle (PBS), IL-1β (5 ng/mL) or IL-1β (5 ng/mL) + IL-1R1 Ab (1 μg/ml) (n = 5 biological replicates per group) (f) for 72 h. Col3a1 mRNA expression was assessed using RT-qPCR. Fold change of Col3a1 expression is shown in bar graphs as mean ± SEM. Statistical significance was determined using one-way ANOVA with Tukey’s multiple comparison test. g Immunofluorescence images showing increased Col3a1 expression in MSCs upon IL-1β (5 ng/mL) and IL-1α (5 ng/mL) stimulation and, IL-1R1 Ab (1 μg/ml) treatment abolished IL-1α/β-induced Col3a1 expression in the BM MSCs. Col3a1 (green) and DAPI (blue); scale bars, 100 µm. Representative images from 3 independent experiments are shown. Source data are provided as a Source Data file.