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. 2023 Aug 14;20(10):1171–1185. doi: 10.1038/s41423-023-01073-2

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© The Author(s), under exclusive licence to CSI and USTC 2023. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

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Fig. 2 — NAMPT function is required for the expression of chemokines and immunomodulatory genes in inflammatory cytokine-primed MSCs. A, B RNA-seq was performed on MSCs stimulated with IFN-γ and TNF-α (10 ng/ml each) for 24 h in the presence or absence of 50 nM FK866. The enrichment of chemotaxis and immune response genes was analyzed by GSEA. NES, normalized enrichment score; Padj, adjusted P value: two-tailed, corrected for multiple comparisons using the Benjamini‒Hochberg method (A). Volcano plot of differentially expressed chemokine and anti-inflammatory mediator genes (B) (n = 3). C–F The expression of chemokines and immunomodulatory genes in MSCs stimulated with IFN-γ and TNF-α (10 ng/ml each) for 24 h in the presence or absence of 50 nM FK866 was assayed by qRT‒PCR (n = 3). G The expression of HO1, COX2, iNOS and β-actin (loading control) in MSCs stimulated with IFN-γ and TNF-α (10 ng/ml each) for 24 h in the presence or absence of 50 nM FK866 were determined by immunoblotting. H MSCs were stimulated with IFN-γ and TNF-α (10 ng/ml each) for 24 h in the presence or absence of 50 nM FK866, and the supernatants was assayed for nitrate by a modified Griess reagent (n = 3). I NAD⁺ levels in MSCs stimulated with IFN-γ and TNF-α (10 ng/ml each) with or without 50 nM FK866, 1 mM NMN or both for 24 h (n = 3). J–M The expression of chemokines and immunomodulatory genes in MSCs stimulated with IFN-γ and TNF-α (10 ng/ml each) with or without 50 nM FK866, 1 mM NMN or both for 24 h were determined by qRT‒PCR (n = 3). N MSCs were stimulated with IFN-γ and TNF-α (10 ng/ml each) for 24 h in the presence or absence of 50 nM FK866 and 1 mM NMN, and the supernatants was assayed for nitrate by a modified Griess reagent (n = 3). O NAD⁺ levels in MSCs stimulated with IFN-γ and TNF-α (5 ng/ml each) for 24 h in the presence or absence of 5 μM P7C3 (n = 3). P–R The expression of immunomodulatory genes in MSCs stimulated with IFN-γ and TNF-α (5 ng/ml each) for 24 h in the presence or absence of 5 μM P7C3 was assayed by qRT‒PCR (n = 3). Values are presented as the mean ± SEM. Statistical analysis was performed by one-way analysis of variance