Fig 2: Lyn facilitated the nuclear translocation of IRF5. (A) Representative images of protein expressions of n‐p‐IRF5, n‐IRF5, cy‐p‐IRF5, cy‐IRF5 after Bafetinib and siRNA‐lyn intrathecally in CCI mice; (B) statistical analysis of protein expressions of n‐p‐IRF5, n‐IRF5, cy‐p‐IRF5, cy‐IRF5 after Bafetinib and siRNA‐lyn intrathecally in CCI mice; (C) representative images of protein expressions of n‐p‐IRF5, n‐IRF5, cy‐p‐IRF5, cy‐IRF5 after Bafetinib and siRNA‐lyn administration in cultured primary microglia; (D) statistical analysis of protein expressions of n‐p‐IRF5, n‐IRF5, cy‐p‐IRF5, cy‐IRF5 after Bafetinib and siRNA‐lyn administration in cultured primary microglia.

Fig 3: IRF5 promoted the binding of transcription factors SP1, PU.1 to glycolytic gene promoters. (A) The binding of SP1 to glycolytic gene promoters HK1, TPI1, PGAM1, PFKFB3 and positive controls IL‐6, IFN‐α after siRNA‐IRF5 intrathecally in CCI mice by ChIP; (B) the binding of PU.1 to glycolytic gene promoters and positive controls after siRNA‐IRF5 intrathecally in CCI mice by ChIP; (C) the binding of SP1 to glycolytic gene promoters and positive controls after siRNA‐IRF5 administration in cultured primary microglia by ChIP; (D) the binding of PU.1 to glycolytic gene promoters and positive controls after siRNA‐IRF5 administration in cultured primary microglia by ChIP.

Fig 4: GEO database reanalysis showed that Lyn might be associated with neuropathic pain. (A) HCA plot of CCI and sham mice in GSE186237 datasets in GEO database; (B) PCA plot of CCI and sham mice; (C) bar graph presented the number of DEGs; (D) the GO analysis results; (E) the KEGG pathway enrichment results; (F) heatmap that showed the DEGs contained in these pathways including Lyn; (G) heatmap that showed the DEGs contained in these pathways including IRF5.

Fig 5: Chronic nerve injury induces the increased expression and phosphorylation of Lyn to mediate IRF5 phosphorylation and nuclear translocation in microglia. Nuclear phosphorylated IRF5 promotes the binding of transcription factors SP1, PU.1 to glycolytic gene promoters and enhances glycolysis of microglia, resulting in the increased production of lactate and other metabolites. Lactate facilitates the proliferation and pro‐inflammatory phenotype transition of microglia to release inflammatory factors such as TNF, IL‐1β, HIF‐1α, PGE2, which contributes vitally to M1 polarization and neruoinflammation. Lactate and other metabolites also facilitates the sEPSC of the postsynaptic neuron. Neruoinflammation and strengthened sEPSC both promote the progression of neuropathic pain. IRF5, interferon regulatory factor 5; TNF, tumour necrosis factor; IL‐1β, interleukin 1β; HIF‐1α, hypoxia inducible factor‐1α; PGE2, prostaglandin E2; sEPSC, spontaneous excitatory postsynaptic currents.