Journal: Arthritis Research & Therapy

Article Title: IRF7 orchestrates proinflammatory macrophage polarization and joint destruction in rheumatoid arthritis

doi: 10.1186/s13075-025-03708-3

Figure Lengend Snippet: Increased frequency and proinflammatory signature of CD48 high S100A12 + macrophages in rheumatoid arthritis synovium. A UMAP visualization of synovial macrophages clustered into nine subpopulations on the basis of scRNA-seq data. B Disease-stratified analysis showing an increased proportion of CD48 high S100A12 + macrophages in the RA synovium compared with those in the UA, OA, and HC groups. C UMAP plots depicting S100A12 expression intensity across macrophage subclusters in different disease states. D Gene Ontology (GO) enrichment of biological processes in CD48 high S100A12 + marker genes, highlighting enrichment for defense response activation, cytokine production, and leukocyte migration. E Representative images of CD68 and S100A12 immunofluorescence staining in knee synovial tissues from RA and OA patients. Scale bar: 100 μm

Article Snippet: Double immunofluorescence staining was performed as follows: sections were incubated overnight at 4 °C with primary antibodies against CD68 (mouse monoclonal, 1:100 dilution; 66231-2-Ig, Proteintech, China) with S100A12 (rabbit polyclonal, 1:100 dilution; 16630-1-AP, Proteintech, China) or CD68 (1:100 dilution; 66231-2-Ig) with IRF7 (rabbit polyclonal, 1:150 dilution; 22392-1-AP, Proteintech, China).

Techniques: Expressing, Marker, Activation Assay, Migration, Immunofluorescence, Staining

Journal: Arthritis Research & Therapy

Article Title: IRF7 orchestrates proinflammatory macrophage polarization and joint destruction in rheumatoid arthritis

doi: 10.1186/s13075-025-03708-3

Figure Lengend Snippet: IRF7 is a specific transcriptional regulator of CD48 high S100A12 + macrophages. A Venn diagram showing overlapping transcription factors (TFs) identified by triplicate SCENIC analyses, with the CD48 high S100A12 + subcluster enriched for NFIL3, TGIF1, FOSL2, IRF7, and STAT1. B Heatmap of regulon activity scores (RASs) for TFs across macrophage subclusters. C Ranking of TFs in CD48 high S100A12 + macrophages by the regulon specificity score (RSS, calculated via Jensen‒Shannon divergence). D UMAP dimensionality reduction of TF activity profiles across subclusters. E – F UMAP plots highlighting spatial overlap between the CD48 high S100A12 + subcluster. ( E ) and cells with elevated IRF7 regulon activity ( F ). G Representative images of immunofluorescence staining for CD68 and IRF7 in knee synovial tissues from RA and OA patients. Scale bar: 100 μm

Article Snippet: Double immunofluorescence staining was performed as follows: sections were incubated overnight at 4 °C with primary antibodies against CD68 (mouse monoclonal, 1:100 dilution; 66231-2-Ig, Proteintech, China) with S100A12 (rabbit polyclonal, 1:100 dilution; 16630-1-AP, Proteintech, China) or CD68 (1:100 dilution; 66231-2-Ig) with IRF7 (rabbit polyclonal, 1:150 dilution; 22392-1-AP, Proteintech, China).

Techniques: Activity Assay, Immunofluorescence, Staining

Journal: Arthritis Research & Therapy

Article Title: IRF7 orchestrates proinflammatory macrophage polarization and joint destruction in rheumatoid arthritis

doi: 10.1186/s13075-025-03708-3

Figure Lengend Snippet: IRF7 directly regulates downstream inflammatory genes in M1 macrophages. A ChIP-seq peak heatmaps showing increased IRF7 binding to promoter/enhancer regions in LPS-stimulated M1 macrophages. B Venn diagram of 108 overlapping genes from the IRF7 ChIP-seq data and the SCENIC-predicted target genes. C Reactome pathway enrichment of IRF7-regulated genes, highlighting the involvement of NF-κB, TNF, and Toll-like receptor signalling (key genes: IL-1β, FOS, NF-κB1, PTGS2, and CXCL10). D Bulk RNA-seq heatmap showing the upregulation of IRF7 and target genes in M1-polarized macrophages ( GSE130011 , GSE154346 ). E UMAP plots of NFKB1, PTGS2, IL1B, and CXCL10 expression in the CD48 high S100A12 + subcluster. F RT‒qPCR analysis of IRF7 and M1 marker genes in siRNA-treated macrophages (performed in triplicate, with 3 distinct patient sources used for each repetition). G – H Western blot validation of IRF7 and downstream protein expression following IRF7 knockdown in M1-polarized macrophages (performed in triplicate, with 3 distinct patient sources used for each repetition). Statistical significance: * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 (one-way ANOVA with the Bonferroni post hoc correction)

Article Snippet: Double immunofluorescence staining was performed as follows: sections were incubated overnight at 4 °C with primary antibodies against CD68 (mouse monoclonal, 1:100 dilution; 66231-2-Ig, Proteintech, China) with S100A12 (rabbit polyclonal, 1:100 dilution; 16630-1-AP, Proteintech, China) or CD68 (1:100 dilution; 66231-2-Ig) with IRF7 (rabbit polyclonal, 1:150 dilution; 22392-1-AP, Proteintech, China).

Techniques: ChIP-sequencing, Binding Assay, RNA Sequencing, Expressing, Marker, Western Blot, Biomarker Discovery, Knockdown

Journal: Arthritis Research & Therapy

Article Title: IRF7 orchestrates proinflammatory macrophage polarization and joint destruction in rheumatoid arthritis

doi: 10.1186/s13075-025-03708-3

Figure Lengend Snippet: Local IRF7 knockdown alters the immune cell composition in CIA mice. A Schematic of intra-articular IRF7 siRNA treatment in collagen-induced arthritis (CIA) model mice. B – C Flow cytometry analysis of the CD86 and CD206 mean fluorescence intensities (MFIs) in F4/80 + macrophages from ankle joints ( n = 6). CD86: NC: 1241 ± 265.4, si-IRF7: 2469 ± 390.3, positive: 3489 ± 570.9, si-mock: 3689 ± 370.1. CD206: NC: 2225 ± 225.9, si-IRF7: 3395 ± 369.4, positive: 978.2 ± 147.9, si-mock: 2022 ± 170.6. D – E Frequencies of Foxp3 + Tregs among CD3 + CD4 + T cells ( n = 6). NC: 1.66% ± 0.15%, si-IRF7: 3.27% ± 0.28%, positive: 0.47% ± 0.17%, si-mock: 1.10% ± 0.22%. F Immunofluorescence staining for S100A12 + inflammatory macrophages in the ankle synovium of different groups. Scale bar: 100 μm. The data are presented as the means ± SDs. Statistical significance: *** P < 0.001, **** P < 0.0001 (one-way ANOVA with Bonferroni post hoc correction)

Article Snippet: Double immunofluorescence staining was performed as follows: sections were incubated overnight at 4 °C with primary antibodies against CD68 (mouse monoclonal, 1:100 dilution; 66231-2-Ig, Proteintech, China) with S100A12 (rabbit polyclonal, 1:100 dilution; 16630-1-AP, Proteintech, China) or CD68 (1:100 dilution; 66231-2-Ig) with IRF7 (rabbit polyclonal, 1:150 dilution; 22392-1-AP, Proteintech, China).

Techniques: Knockdown, Flow Cytometry, Fluorescence, Immunofluorescence, Staining

Journal: Arthritis Research & Therapy

Article Title: IRF7 orchestrates proinflammatory macrophage polarization and joint destruction in rheumatoid arthritis

doi: 10.1186/s13075-025-03708-3

Figure Lengend Snippet: Local IRF7 inhibition attenuates joint inflammation and bone erosion in CIA mice. A Representative ankle joint images on day 42 postimmunization. B H&E staining and histological staining. C - E IHC staining for CD68, S100A12, and IRF7 in the ankle synovium. Scale bar: 100 μm. F Quantification of paw thickness at the ankle joint ( n = 6 per group). 42 Days after the first immunization: NC: 8.33 ± 0.02, si-IRF7: 10.39 ± 0.54, positive: 12.08 ± 0.80, si-mock: 12.65 ± 0.57, Statistical significance: **** P < 0.0001 (one-way ANOVA with Bonferroni post hoc correction). G H&E staining and histological scoring of synovial hyperplasia and inflammation ( n = 6). NC: 0.00 (0.00–0.00), si-IRF7: 1.50 (1.00–2.25), positive: 2.50 (1.75–3.00), and si-mock: 3.00 (2.75–3.00). Data are shown as medians with 25% − 75% percentiles. Statistical significance: * P < 0.05 (Kruskal‒Wallis test, followed by post hoc Dunn’s test with Bonferroni correction for multiple comparisons). H - J Semiquantitative analysis analysis of CD68, S100A12, and IRF7 expression via IHC staining via ImageJ ( n = 6). Statistical significance: **** P < 0.0001 (one-way ANOVA with Bonferroni post hoc correction)

Article Snippet: Double immunofluorescence staining was performed as follows: sections were incubated overnight at 4 °C with primary antibodies against CD68 (mouse monoclonal, 1:100 dilution; 66231-2-Ig, Proteintech, China) with S100A12 (rabbit polyclonal, 1:100 dilution; 16630-1-AP, Proteintech, China) or CD68 (1:100 dilution; 66231-2-Ig) with IRF7 (rabbit polyclonal, 1:150 dilution; 22392-1-AP, Proteintech, China).

Techniques: Inhibition, Staining, Immunohistochemistry, Expressing

Journal: PLoS ONE

Article Title: Inhibition of Neuronal Apoptosis and Axonal Regression Ameliorates Sympathetic Atrophy and Hemodynamic Alterations in Portal Hypertensive Rats

doi: 10.1371/journal.pone.0084374

Figure Lengend Snippet: Bar diagram showing CGRP quantitation by Western blot analysis in sham (n = 6) and PVL (n = 7). Representative Western blot is shown below.

Article Snippet: Protein expression analysis was performed in nodose ganglion samples for calcitonin gene related protein (CGRP) (diluted 1/500) (Biomol, Pennsylvania PA, USA) and in SMG for tyrosine hydroxylase (Th) (dil 1/600), Sema 3A, sortilin (Sort1) (diluted 1/1000) (Abcam, Cambridge, UK), TrkA, Rho kinase (ROCK) (diluted 1/200) (Santa Cruz Biotechnology, Santa Cruz, CA, USA), phospho-TrkA (pTrkA), mitogen activated protein kinase (MAPK), phospho-MAPK (pMAPK), c-Jun N-terminal kinase (JNK), phospho-JNK (pJNK), protein kinase B (AKT), phospho-AKT (pAKT), cleaved caspase7 (casp-7), poly(ADP-ribose) polymerase (PARP) (diluted 1/1000) (Cell Signaling Technology, Beverly, MA, USA) and neural nitric oxide synthase (nNOS) (1/1000) (BD Biosciences, San Jose, CA, USA).

Techniques: Quantitation Assay, Western Blot