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. 2026 Aug 5;12(32):eaea2780. doi: 10.1126/sciadv.aea2780

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Copyright © 2026 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works. Distributed under a Creative Commons Attribution NonCommercial License 4.0 (CC BY-NC).

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Fig. 2. — (A) Venn diagrams of DEGs in parental and N-expressing THP-1 macrophage-like cells after PRR stimulation. DEGs were defined as FDR < 0.05 (stimulated versus untreated), regardless of log₂ fold change. DEGs were classified as N-specific, parental-specific, or shared. CL097 (TLR7/8) altered 1974 shared and 1422 N-specific genes; poly(I:C) (TLR3) altered 1583 shared and 2827 N-specific genes; poly(I:C) transfection (RIG-I/MDA5) altered 3691 shared and 1979 N-specific genes. (B) Histograms showing N-induced changes in DEG expression, represented as delLog₂FC distributions for each PRR stimulation condition. delLog₂FC was calculated by comparing log₂ fold changes in N-expressing cells to parental cells under the same stimulation condition. (C) BP enrichment analysis of N-induced DEGs. Left: Top 10 GO terms from 69 N–up-regulated DEGs after TLR7/8 stimulation. Middle: Top 10 GO terms from 140 N–up-regulated DEGs after TLR3 stimulation. Right: Top 10 GO terms from 144 N–down-regulated DEGs after RIG-I/MDA5 stimulation. Bars above the −log₁₀(FDR) cutoff (red line, FDR = 0.05) indicate significant enrichment. (D to F) Fold change bar graphs of representative DEGs dramatically regulated by N in PRR-stimulated THP-1 macrophage-like cells. Log₂ fold changes of DEGs exhibiting enhanced expression in both parental and N-expressing cells to different extents, and those selectively up-regulated in N-expressing cells after TLR7/8 stimulation (D), TLR3 stimulation (E), or RIG-I/MDA5 stimulation (F). (G) Heatmap of innate immune response DEGs showing N-induced expression changes across PRR stimulations. DEGs from parental and N-expressing THP-1 macrophage-like cells were filtered using innate immune BP and Kyoto Encyclopedia of Genes and Genomes (KEGG) terms (GO:0140374, GO:0051607, GO:0009615, GO:0032728, GO:0070106, GO:0006954, GO:0045071, GO:0050729, GO:0045087, GO:0060337, GO:0032760, GO:0050728). Colors represent delLog₂FC [N versus WT log₂FC under the same PRR condition; see (B)]. Red, up-regulated by N; blue, down-regulated by N.