Journal: Frontiers in Immunology

Article Title: A20 mRNA therapeutics ameliorate systemic sclerosis by suppressing TRAF-6/NF-KB signaling and DREAM expression and exerting antifibrotic effects

doi: 10.3389/fimmu.2025.1665998

Figure Lengend Snippet: A20 mRNA Therapy Suppressed the Expression of DREAM, a Known A20 Inhibitory Molecule, and Reduced SMAD2, αSMA, and Procollagen in Systemic Sclerosis Skin Tissue and Human Skin Fibroblasts. (A) Skin and lung sections were stained with antibodies against DREAM, SMAD2, procollagen, and αSMA. The number of antibody-positive cells (mean ± SEM) is plotted. Original magnification: 400×; scale bar: 100 μm. (B) The protein levels of DREAM and SMAD2 were measured in cells treated with TGF-β (20 ng/mL) for 48 h GAPDH served as the validation control. For quantification, three fields per mouse were analyzed (n = 5 mice per group). Data are presented as mean ± SD from a representative experiment of three independent experiments. Statistical significance was determined by one-way ANOVA (A, B) . (*P < 0.05; **P < 0.01).

Article Snippet: Antibodies targeting A20 (ab13597, Abcam, Cambridge, UK), TRAF6 (SC-8409, Santa Cruz Technologies, Santa Cruz, CA, USA), NF-κB (ab16502, Abcam), DREAM (SC-166916, Santa Cruz Technologies), SMAD2 (ab33875, Abcam), Col1a1 (PA5-29569, Invitrogen, CA, USA) αSMA (ab7817, Abcam), c-Myc (SC-40, Santa Cruz), p53 (SC-6243, Santa Cruz) and GAPDH (ab181602, Abcam) were used to measure the corresponding protein levels by Western blotting (SNAP i.d.

Techniques: Expressing, Staining, Biomarker Discovery, Control

Journal: Frontiers in Immunology

Article Title: A20 mRNA therapeutics ameliorate systemic sclerosis by suppressing TRAF-6/NF-KB signaling and DREAM expression and exerting antifibrotic effects

doi: 10.3389/fimmu.2025.1665998

Figure Lengend Snippet: A20 mRNA Therapy Reduces Fibrosis in Systemic Sclerosis Lung Tissue by Inhibiting DREAM Expression. Lung sections were stained with antibodies against DREAM, SMAD2, procollagen, and αSMA. For quantification, three fields per mouse were analyzed (n = 5 mice per group), and the number of antibody-positive cells is plotted as mean ± SEM. Original magnification: 400×; scale bar: 100 μm. Bar graph shows data from one representative experiment (**P < 0.01). The statistical significance of all graphs was determined by one-way ANOVA (A) . The English in this document has been checked by at least two professional editors, both native speakers of English. For a certificate, please see: http://www.textcheck.com/certificate/bAoEK8 .

Article Snippet: Antibodies targeting A20 (ab13597, Abcam, Cambridge, UK), TRAF6 (SC-8409, Santa Cruz Technologies, Santa Cruz, CA, USA), NF-κB (ab16502, Abcam), DREAM (SC-166916, Santa Cruz Technologies), SMAD2 (ab33875, Abcam), Col1a1 (PA5-29569, Invitrogen, CA, USA) αSMA (ab7817, Abcam), c-Myc (SC-40, Santa Cruz), p53 (SC-6243, Santa Cruz) and GAPDH (ab181602, Abcam) were used to measure the corresponding protein levels by Western blotting (SNAP i.d.

Techniques: Expressing, Staining

Journal: Frontiers in Immunology

Article Title: A20 mRNA therapeutics ameliorate systemic sclerosis by suppressing TRAF-6/NF-KB signaling and DREAM expression and exerting antifibrotic effects

doi: 10.3389/fimmu.2025.1665998

Figure Lengend Snippet: Proposed mechanism by which A20 mRNA therapy attenuates fibrosis in systemic sclerosis. In the systemic sclerosis condition (left), activation of TLR4 initiates TRAF6-mediated NF-κB signaling and upregulates TGF-β, IL-8, collagen type I (Col1), and α-smooth muscle actin (α-SMA), thereby promoting fibroblast activation and extracellular matrix (ECM) accumulation. In contrast, A20 mRNA encapsulated in lipid nanoparticles (LNPs) (right) restores A20 protein expression, leading to inhibition of TRAF6-dependent NF-κB phosphorylation, suppression of DREAM and SMAD pathways, and subsequent reduction of fibrotic markers in skin and lung fibroblasts. Downward arrows indicate decreased expression following A20 mRNA treatment. The schematic was created with BioRender.com .

Article Snippet: Antibodies targeting A20 (ab13597, Abcam, Cambridge, UK), TRAF6 (SC-8409, Santa Cruz Technologies, Santa Cruz, CA, USA), NF-κB (ab16502, Abcam), DREAM (SC-166916, Santa Cruz Technologies), SMAD2 (ab33875, Abcam), Col1a1 (PA5-29569, Invitrogen, CA, USA) αSMA (ab7817, Abcam), c-Myc (SC-40, Santa Cruz), p53 (SC-6243, Santa Cruz) and GAPDH (ab181602, Abcam) were used to measure the corresponding protein levels by Western blotting (SNAP i.d.

Techniques: Activation Assay, Expressing, Inhibition, Phospho-proteomics