Journal: Journal of Translational Autoimmunity

Article Title: A novel epigenetic regulation of JAM-A by EZH2-DNMT3A cascade contributes to T cell adhesion via the activation of Rap1a in lupus patients

doi: 10.1016/j.jtauto.2026.100362

Figure Lengend Snippet: Increased expression of JAM-A in SLE patient PBMCs and peripheral T cells. (A – D) FCM analysis of JAM-A + PBMCs and JAM-A + T cells. (A) JAM-A was stained with anti-human JAM-A-PE antibodies in the PBMCs of healthy individuals (red), new SLE patients (blue), SLE treated (green) and isotype controls (black). (B) Quantification of JAM-A + PBMCs from (A). (C) Double staining with both anti-human JAM-A-PE and anti-human CD3-FITC antibodies. (D) Quantification of JAM-A + T cells from (C), and the error bars indicate the SEM. Healthy, (healthy volunteers, n = 24); new SLE (SLE patients without clinical treatments, n = 9); SLE treated (SLE patients with clinical treatments, n = 15). (E – F) The correlations of EZH2 mRNA/β-actin and the JAM-A mRNA/β-actin in PBMCs ( E ) and T cells ( F ) from SLE patients. ∗ P < 0.05, ∗∗∗ P < 0.001.

Article Snippet: Cells were stimulated overnight with anti-CD3 and anti-CD28 antibodies and transfected with 100 nM EZH2 siRNAs (siG09121182926-1-5/si-h-EZH2_001 and siG09121182954-1-5/si-h-EZH2_002, RiboBio, China), JAM-A siRNAs (SC-43139, Santa Cruz Biotechnology, USA), and negative control siRNA (siN0000001-1-5 for EZH2, SC-37007 for JAM-A) in 6-well plates.

Techniques: Expressing, Staining, Double Staining

Journal: Journal of Translational Autoimmunity

Article Title: A novel epigenetic regulation of JAM-A by EZH2-DNMT3A cascade contributes to T cell adhesion via the activation of Rap1a in lupus patients

doi: 10.1016/j.jtauto.2026.100362

Figure Lengend Snippet: The crosstalk between EZH2/H3K27me3 and DNMT3A/JAM-A. (A, D) The protein expression in Jurkat cells or T cells treated with 5 μM GSK126 (A) or EZH2-siRNA (D), GAPDH or H3 protein as loading control. (B, E) The methylation frequency in JAM-A gene promoter of T cells under EZH2 inhibitor 5 μM GSK126 (B) or EZH2-siRNA treated (E). (C, F) Immunoprecipitated JAM-A with anti-DNMT3A or DNMT3A with anti-H3K27me3 ChIP-grade antibodies in T cells under 5 μM GSK126 (C) or EZH2-siRNA treated (F). The values were normalized to the chromatin levels with 1% input samples. (G, H) The negative correlations of EZH2 mRNA/β-actin and DNMT3A mRNA/β-actin (G) , DNMT3A mRNA/β-actin and JAM-A mRNA/β-actin (H) in T cells from SLE patients. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001, and error bars indicate the SEM.

Article Snippet: Cells were stimulated overnight with anti-CD3 and anti-CD28 antibodies and transfected with 100 nM EZH2 siRNAs (siG09121182926-1-5/si-h-EZH2_001 and siG09121182954-1-5/si-h-EZH2_002, RiboBio, China), JAM-A siRNAs (SC-43139, Santa Cruz Biotechnology, USA), and negative control siRNA (siN0000001-1-5 for EZH2, SC-37007 for JAM-A) in 6-well plates.

Techniques: Expressing, Control, Methylation, Immunoprecipitation

Journal: Journal of Translational Autoimmunity

Article Title: A novel epigenetic regulation of JAM-A by EZH2-DNMT3A cascade contributes to T cell adhesion via the activation of Rap1a in lupus patients

doi: 10.1016/j.jtauto.2026.100362

Figure Lengend Snippet: Inhibition of EZH2 and JAM-A suppresses β1 integrin-mediated T cells and Jurkat cells adhering to ECM. ( A ) The adhesion capacity in T cells from Healthy and SLE patients in vitro . ( B-C ) The adhesion capacity of T cells isolated from healthy or Jurkat cells blocked by 5 μM GSK126 (B) or EZH2-siRNA (C) in vitro . ( D ) Compared the adhesion capacity of T cells from SLE patients, or Jurkat cells with/out neutralizing anti-JAM-A, 1 μg/ml J10.4. (E) Protein expression levels in Jurkat cells overexpressing or knockdown of JAM-A. (F) The adhesion capacity of Jurkat cells with JAM-A regulation. ∗P < 0.05, ∗∗P < 0.01, ∗∗∗ P < 0.001, and error bars indicate the SEM.

Article Snippet: Cells were stimulated overnight with anti-CD3 and anti-CD28 antibodies and transfected with 100 nM EZH2 siRNAs (siG09121182926-1-5/si-h-EZH2_001 and siG09121182954-1-5/si-h-EZH2_002, RiboBio, China), JAM-A siRNAs (SC-43139, Santa Cruz Biotechnology, USA), and negative control siRNA (siN0000001-1-5 for EZH2, SC-37007 for JAM-A) in 6-well plates.

Techniques: Inhibition, In Vitro, Isolation, Expressing, Knockdown

Journal: Journal of Translational Autoimmunity

Article Title: A novel epigenetic regulation of JAM-A by EZH2-DNMT3A cascade contributes to T cell adhesion via the activation of Rap1a in lupus patients

doi: 10.1016/j.jtauto.2026.100362

Figure Lengend Snippet: Inhibition of EZH2 with GSK126 ameliorates lupus-like disease phenotypes in MRL/ lpr mice. (A) Eight-week-old female MRL/ lpr mice were adaptively fed for 2 weeks (wks, n = 15) in the SPF animal room. Mice were randomly divided into a vehicle group (n = 8) and a GSK126-treated group (n = 7) and were treated with an intraperitoneal (IP) injection of β-SEB (20%) or GSK126 (50 mg/kg) every other day for one month. (B) Survival curves of the vehicle and treated group. (C) Anti-dsDNA antibody levels in the plasma. (D) The mRNA expression of IL-10 and TGF-β in splenomegaly. (E) Photomicrographic representation of renal damage (arrow). (F) The percentage plot of glomerulus damage. (G) The Representative histological images of immunohistochemical staining of JAM-A and β1-integrin antibody. (H) The quantification of positive staining areas was quantified by Saiviewer software. (I) The protein expression of EZH2, DNMT3A, JAM-A, Rap1a, β1 integrin and H3K27me3 in splenocytes detected by western blotting. (J) A total of 7 mice per group were analyzed for densitometry by image J software from (I). The symbols represent individual mice, and error bars indicate the SEM. ∗P < 0.05, ∗∗∗P < 0.001, ∗∗∗P < 0.001.

Article Snippet: Cells were stimulated overnight with anti-CD3 and anti-CD28 antibodies and transfected with 100 nM EZH2 siRNAs (siG09121182926-1-5/si-h-EZH2_001 and siG09121182954-1-5/si-h-EZH2_002, RiboBio, China), JAM-A siRNAs (SC-43139, Santa Cruz Biotechnology, USA), and negative control siRNA (siN0000001-1-5 for EZH2, SC-37007 for JAM-A) in 6-well plates.

Techniques: Inhibition, Injection, Clinical Proteomics, Expressing, Immunohistochemical staining, Staining, Software, Western Blot

Journal: Journal of Translational Autoimmunity

Article Title: A novel epigenetic regulation of JAM-A by EZH2-DNMT3A cascade contributes to T cell adhesion via the activation of Rap1a in lupus patients

doi: 10.1016/j.jtauto.2026.100362

Figure Lengend Snippet: Proposed schematic model of the EZH2/miR-26a-5p signaling axis involved in the development of SLE. Increased EZH2 expression is mediated by miR-26a-5p, which in turn is epigenetically repressed by EZH2-mediated H3K27me3 trimethylation within the miR-26a-5p promoter, thus forming a vicious cycle. Crosstalk between H3K27me3 and CpG island methylation mediated by DNMT3A within the JAM-A promoter, resulted in increased expression of JAM-A. Increased expression of JAM-A up-regulated the expression of Rap1a, a regulator of β1-integrin, which is functionally relevant to T cell adhesion. Combined with the increased transcriptional level of Rap1a mediated by miR-26a-5p, ultimately, Rap1a led to increased expression of β1-integrin, which is involved in increasing T cell adhesion.

Article Snippet: Cells were stimulated overnight with anti-CD3 and anti-CD28 antibodies and transfected with 100 nM EZH2 siRNAs (siG09121182926-1-5/si-h-EZH2_001 and siG09121182954-1-5/si-h-EZH2_002, RiboBio, China), JAM-A siRNAs (SC-43139, Santa Cruz Biotechnology, USA), and negative control siRNA (siN0000001-1-5 for EZH2, SC-37007 for JAM-A) in 6-well plates.

Techniques: Expressing, Methylation