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. 2025 Jul 31;10(8):e00361-25. doi: 10.1128/msphere.00361-25

Fig 3.

Heatmap, bar charts, and immunoblots depict reduced expression and activation of NF kappa B pathway targets in NMN-treated group, consistent with downregulated cytokine signaling and altered nuclear localization of transcription factor p65.

NMN reduces the inflammatory response of macrophages infected with hvKP. Marrow-derived macrophages (BMDMs) were treated with NMN (500 µM) for 16 h and infected with ATCC 43816 for 3 h (MOI = 1). (A) Hierarchical clustering heatmap of selected significant differentially expressed genes (DEGs) in BMDMs from NMN-treated and untreated controls. Red denotes increased expression; blue denotes decreased expression. (B) mRNA levels of Il6, Il1b, Tnf, and Cxcl2 between NMN-treated and untreated controls. (C) Downregulated KEGG pathway enrichment after treatment with NMN. (D) GSEA analysis of NF-κB signaling pathway. (E) Western blot analysis of p-p65, p65, p-IκBα, and IκBα protein expression levels among four groups. (F) Immunofluorescence analysis of colocalization between p65 and nucleus (Hoechst 33258). Data are representative results from three independent experiments (mean  ±  SEM). *, P <  0.05; **, P <  0.01; ***, P <  0.001; ****, P < 0.0001; ns, not significant. NMN, β-nicotinamide mononucleotide; BMDM, bone marrow-derived macrophage; hvKP, hypervirulent Klebsiella pneumoniae; MOI, multiplicity of infection; Control, no treatment or infection group.