Journal: Journal of Translational Autoimmunity

Article Title: Exploring the immunomodulatory effects of environmental contaminants on autoimmune patients: An in vitro approach

doi: 10.1016/j.jtauto.2025.100341

Figure Lengend Snippet: Modulation of intracellular signaling pathways in HD, RA, and SLE cells exposed to environmental contaminants. Heatmap showing the relative expression and phosphorylation levels of key signaling proteins involved in immune regulation, inflammation, and cell survival (STAT1, STAT3, p38, AKT, and NFκB) and their phosphorylated forms in HD, RA, and SLE after exposure to PM, silica, and TCDD. Data are represented as z-score normalized values. Asterisks indicate statistically significant differences compared with the unstimulated control within each group (p < 0.05). AKT: protein kinase B; HD: healthy donors; NFκB: nuclear factor kappa-light-chain-enhancer of activated B cells; P: phosphorylated form; p38: p38 mitogen-activated protein kinase; PM: particulate matter; RA: rheumatoid arthritis; SLE: systemic lupus erythematosus; STAT: signal transducer and activator of transcription; TCDD: 2,3,7,8-tetrachlorodibenzo-p-dioxin.

Article Snippet: Membranes were blocked with 5 % non-fat milk in TBS-T and incubated overnight at 4 °C with primary antibodies against phospho-AKT (Thr308), phospho-NFκB p65 (Ser536), phospho-p38 MAPK (Thr180/Tyr182), phospho-STAT1 (Tyr701), and phospho-STAT3 (Tyr705) (Cell Signaling Technology, Danvers, MA, USA; Santa Cruz Biotechnology, Dallas, TX, USA).

Techniques: Protein-Protein interactions, Expressing, Phospho-proteomics, Control

Journal: Genes & Diseases

Article Title: MFAP2 promotes metastasis and drug resistance by regulating epithelial-to-mesenchymal transition through EGFR signaling pathway in colorectal cancer cells

doi: 10.1016/j.gendis.2025.101800

Figure Lengend Snippet: MFAP2 promotes epithelial–mesenchymal transition (EMT) through the EGFR-AKT-STAT3 signaling pathway in colorectal cancer (CRC) cells. (A, B) The top 20 enrichment signaling pathways regulated by MFAP2 knockdown. (C, D) VEGFR2 signaling pathway was enriched in the MFAP2 knockdown cells, shown by Gene Set Enrichment Analysis (GSEA) and essential genes in this enrichment. (E) MFAP2 knockdown affected the EGFR-AKT-STAT3 axis.

Article Snippet: Following blocking with 5% non-fat milk in PBS with 0.02% Tween 20 detergent (PBST) at room temperature for 2 h, the membranes were incubated with primary antibodies, including MFAP2 (Solarbio, China), GAPDH (BBI Co., Ltd., China), epidermal growth factor receptor (EGFR; Proteintech, China), protein kinase B (AKT) (Proteintech), signal transducer and activator of transcription 3 (STAT3) (Proteintech), and vascular endothelial growth factor A (VEGFA) (Proteintech), p-EGFR (Cell Signaling Technology, USA), p-STAT3 (Cell Signaling Technology), and p-AKT ser473 (Cell Signaling Technology) antibodies, at 4 °C overnight.

Techniques: Protein-Protein interactions, Knockdown

Journal: Molecular Medicine Reports

Article Title: Long non-coding RNA NKILA regulates the JAK2/STAT3 pathway to exacerbate TGF-β1-mediated renal fibrosis

doi: 10.3892/mmr.2026.13839

Figure Lengend Snippet: Heat map analysis of the top 20 cis-regulated lncRNAs in the lncRNA high-throughput sequencing and the binding and site results of the NKILA promoter region predicted by JASPAR database. (A) Heatmap demonstrating the top 20 cis-regulated lncRNAs in the control group compared with the normal group. ‘Normal’ indicates blank groups and ‘control’ indicates TGF-β1 (10 ng/ml) induced HK-2 cells. Y-axis shows cis-regulated lncRNA corresponding names. Color changes represent the expression level of the control group changes after normalization treatment compared with the blank group. The change from blue to red after normalization ranges from 0–2. (B) STAT3 binding site information predicted by the JASPAR database for the promoter region from 2000-99 bp upstream of NKILA. lncRNA, long non-coding RNA.

Article Snippet: After blocking with 5% non-fat milk for 2 h at room temperature, membranes were incubated overnight at 4°C with primary antibodies against fibronectin (FN; 1:1,000; cat. no. ab45688; Abcam), collagen I (Col1; 1:1,000; cat. no. ab138492; Abcam), vimentin (Vim; 1:1,000; cat. no. 10366-1-AP; Proteintech Group, Inc.), α-smooth muscle actin (α-SMA; 1:1,000; cat. no. 14395-1-AP; Proteintech Group, Inc.), JAK2 (1:1,000; cat. no. 17670-1-AP; Proteintech Group, Inc.), STAT3 (1:1,000; cat. no. 10253-2-AP; Proteintech Group, Inc.), phosphorylated (p)-JAK2 (1:500; cat. no. ab32101; Abcam), p-STAT3 (1:500; cat. no. ab76315; Abcam) and GAPDH (1:5,000; cat. no. 10494-1-AP; Proteintech Group, Inc.).

Techniques: Next-Generation Sequencing, Binding Assay, Control, Expressing

Journal: Molecular Medicine Reports

Article Title: Long non-coding RNA NKILA regulates the JAK2/STAT3 pathway to exacerbate TGF-β1-mediated renal fibrosis

doi: 10.3892/mmr.2026.13839

Figure Lengend Snippet: JKA2/STAT3 pathway protein detection in HK-2 cells transfected with LncRNA NKILA overexpression virus (A) Representative western blotting bands and (B) semi-quantification of results of protein expression of phosphorylated JAK2 and STAT3 (n=3). (C) RT-qPCR statistical results (n=3). ## P<0.01 and # P<0.05 compared with the normal group. ns, not significant; p-, phosphorylated; OE, overexpression; NC, negative control; JAK, Janus kinase; RT-qPCR, reverse transcription quantitative PCR.

Article Snippet: After blocking with 5% non-fat milk for 2 h at room temperature, membranes were incubated overnight at 4°C with primary antibodies against fibronectin (FN; 1:1,000; cat. no. ab45688; Abcam), collagen I (Col1; 1:1,000; cat. no. ab138492; Abcam), vimentin (Vim; 1:1,000; cat. no. 10366-1-AP; Proteintech Group, Inc.), α-smooth muscle actin (α-SMA; 1:1,000; cat. no. 14395-1-AP; Proteintech Group, Inc.), JAK2 (1:1,000; cat. no. 17670-1-AP; Proteintech Group, Inc.), STAT3 (1:1,000; cat. no. 10253-2-AP; Proteintech Group, Inc.), phosphorylated (p)-JAK2 (1:500; cat. no. ab32101; Abcam), p-STAT3 (1:500; cat. no. ab76315; Abcam) and GAPDH (1:5,000; cat. no. 10494-1-AP; Proteintech Group, Inc.).

Techniques: Transfection, Over Expression, Virus, Western Blot, Expressing, Quantitative RT-PCR, Negative Control, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Molecular Medicine Reports

Article Title: Long non-coding RNA NKILA regulates the JAK2/STAT3 pathway to exacerbate TGF-β1-mediated renal fibrosis

doi: 10.3892/mmr.2026.13839

Figure Lengend Snippet: JAK2/STAT3 signaling pathway-associated proteins in HK-2 cells transfected with LncRNA NKILA knockdown lentivirus (A) Representative western blotting bands and (B) semi-quantification of JAK2/STAT3 pathway protein expression (n=3). (C) Statistical analysis results of reverse transcription-quantitative PCR for channel indicators. ## P<0.01 compared with the normal group (n=3), **P<0.01 and *P<0.05 compared with the control + KD-NC group. JAK, Janus kinase; KD, knockdown; NC, negative control; p-, phosphorylated.

Article Snippet: After blocking with 5% non-fat milk for 2 h at room temperature, membranes were incubated overnight at 4°C with primary antibodies against fibronectin (FN; 1:1,000; cat. no. ab45688; Abcam), collagen I (Col1; 1:1,000; cat. no. ab138492; Abcam), vimentin (Vim; 1:1,000; cat. no. 10366-1-AP; Proteintech Group, Inc.), α-smooth muscle actin (α-SMA; 1:1,000; cat. no. 14395-1-AP; Proteintech Group, Inc.), JAK2 (1:1,000; cat. no. 17670-1-AP; Proteintech Group, Inc.), STAT3 (1:1,000; cat. no. 10253-2-AP; Proteintech Group, Inc.), phosphorylated (p)-JAK2 (1:500; cat. no. ab32101; Abcam), p-STAT3 (1:500; cat. no. ab76315; Abcam) and GAPDH (1:5,000; cat. no. 10494-1-AP; Proteintech Group, Inc.).

Techniques: Transfection, Knockdown, Western Blot, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Control, Negative Control

Journal: Molecular Medicine Reports

Article Title: Long non-coding RNA NKILA regulates the JAK2/STAT3 pathway to exacerbate TGF-β1-mediated renal fibrosis

doi: 10.3892/mmr.2026.13839

Figure Lengend Snippet: JAK2/STAT3 pathway protein detection in the recovery experiment of HK-2 cells treated with AG490. (A) Representative western blotting bands and (B) semi-quantification of JAK2/STAT3 pathway protein expression (n=3). (C) Reverse transcription-quantitative PCR for channel indicators (n=3). ## P<0.01 compared with the normal group, **P<0.01 and *P<0.05 compared with control + DMSO group and △△ P<0.01 compared with OE-NKILA group. OE, overexpression; p-, phosphorylated; JAK, Janus kinase; ns, not significant.

Article Snippet: After blocking with 5% non-fat milk for 2 h at room temperature, membranes were incubated overnight at 4°C with primary antibodies against fibronectin (FN; 1:1,000; cat. no. ab45688; Abcam), collagen I (Col1; 1:1,000; cat. no. ab138492; Abcam), vimentin (Vim; 1:1,000; cat. no. 10366-1-AP; Proteintech Group, Inc.), α-smooth muscle actin (α-SMA; 1:1,000; cat. no. 14395-1-AP; Proteintech Group, Inc.), JAK2 (1:1,000; cat. no. 17670-1-AP; Proteintech Group, Inc.), STAT3 (1:1,000; cat. no. 10253-2-AP; Proteintech Group, Inc.), phosphorylated (p)-JAK2 (1:500; cat. no. ab32101; Abcam), p-STAT3 (1:500; cat. no. ab76315; Abcam) and GAPDH (1:5,000; cat. no. 10494-1-AP; Proteintech Group, Inc.).

Techniques: Western Blot, Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Control, Over Expression

Journal: Frontiers in Aging Neuroscience

Article Title: Unraveling the anti-neuroinflammatory mechanisms of Cervus cucumis polypeptide injection in Alzheimer’s disease: insights from network pharmacology, molecular docking, molecular dynamics simulation, and experimental validation

doi: 10.3389/fnagi.2026.1797302

Figure Lengend Snippet: Effects of CCPI on the expression of IL-6/STAT3/VEGF signaling pathway. (A) Western blot showed the expression of the IL-6/STAT3/VEGF pathway after CCPI treatment. (B–D) Quantitative analysis of the IL-6/GAPDH, STAT3/GAPDH, and VEGF/GAPDH ratios in the CCPI-treated groups, respectively. Data are presented as mean ± SD ( n = 3). ### p < 0.001 vs. ontrol group; *** p < 0.001, ** p < 0.01 and * p < 0.05 vs. AD model group.

Article Snippet: After being blocked in 5% skim milk for 2 h at room temperature, the membranes were incubated overnight with the following primary antibodies: inducible nitric oxide synthase (iNOS) mouse antibody (CAS No. IC259554, Abmart, China, 1:1,000), CD206 mouse antibody (CAS No. ZY-5843R, Abmart, China, 1:1,000), IL-6 mouse antibody (CAS No. 66146-2, Abmart, China, 1:1,000), STAT3 mouse antibody (CAS No. YA056, Abmart, China, 1:1,000), STAT3 phosphorylation mouse antibody (CAS No. 05-485, Sigma, United States, 1:1,000), VEGF rabbit antibody (CAS No. AF1309, Abmart, China, 1:1,000) and GAPDH rabbit antibody (CAS No. 10494-1-AP, Abmart, China, 1:1,000).

Techniques: Expressing, Western Blot

Journal: Frontiers in Aging Neuroscience

Article Title: Unraveling the anti-neuroinflammatory mechanisms of Cervus cucumis polypeptide injection in Alzheimer’s disease: insights from network pharmacology, molecular docking, molecular dynamics simulation, and experimental validation

doi: 10.3389/fnagi.2026.1797302

Figure Lengend Snippet: Comparison of the effects of CCPI and LA on IL-6 secretion, STAT3 phosphorylation, and the expression of markers associated with pro-inflammation (iNOS) and anti-inflammation/repair (CD206) in AD model cells. (A) IL-6 levels in the AD model cells were measured by ELISA. (B) Western blot showed the expression of iNOS, CD206, STAT3 phosphorylation, and STAT3 after CCPI and LA treatment. (C–E) Quantitative analysis of the iNOS/GAPDH, CD206/GAPDH, and STAT3 phosphorylation/STAT3 ratios in the CCPI and LA-treated groups, respectively. Data are presented as mean ± SD ( n = 3). ### p < 0.001 vs. control group; *** p < 0.001, ** p < 0.01 and * p < 0.05 vs. AD model group; ns p > 0.05 compared with the CCPI group; compared with the LA group, ns p > 0.05.

Article Snippet: After being blocked in 5% skim milk for 2 h at room temperature, the membranes were incubated overnight with the following primary antibodies: inducible nitric oxide synthase (iNOS) mouse antibody (CAS No. IC259554, Abmart, China, 1:1,000), CD206 mouse antibody (CAS No. ZY-5843R, Abmart, China, 1:1,000), IL-6 mouse antibody (CAS No. 66146-2, Abmart, China, 1:1,000), STAT3 mouse antibody (CAS No. YA056, Abmart, China, 1:1,000), STAT3 phosphorylation mouse antibody (CAS No. 05-485, Sigma, United States, 1:1,000), VEGF rabbit antibody (CAS No. AF1309, Abmart, China, 1:1,000) and GAPDH rabbit antibody (CAS No. 10494-1-AP, Abmart, China, 1:1,000).

Techniques: Comparison, Phospho-proteomics, Expressing, Enzyme-linked Immunosorbent Assay, Western Blot, Control

Journal: The FASEB Journal

Article Title: CSPG4 Mediates Inflammatory, Cell Death, and Senescence Responses in Enteric Glia Exposed to Clostridioides difficile Toxins

doi: 10.1096/fj.202600333R

Figure Lengend Snippet: CSPG4 blockade reduces TcdB‐induced pSTAT3 nuclear translocation in enteric glial cells. (A) Representative photomicrographs of pSTAT3 (green) immunostaining, phalloidin (red) as a cytoplasm staining, and DAPI (blue) as a nuclear staining in enteric glial cells exposed to TcdA (50 ng/mL) or TcdB (1 ng/mL) at 18 h incubation in the presence of isotype control or anti‐CSPG4 antibodies (0.001 μg/mL). Scale bars: 50 μm. (B) Percentage of enteric glial cells ( n = 5, mean ± SEM) with positive nuclear pSTAT3 staining at 18 h of incubation with TcdA and TcdB in the presence of isotype control or anti‐CSPG4 antibodies (0.001 μg/mL). One‐way ANOVA followed by the Tukey test.

Article Snippet: After washing, cells were incubated overnight at 4°C with primary antibodies against CSPG4 (Abcam, ab275024, 1:400), nectin‐3 (PVRL3, Invitrogen, PA5‐51095, 1:200), LRP1 (Abcam, ab92544, 1:100), NFκB p65 (Santa Cruz Biotechnology, sc‐372, 1:100), or pSTAT3 (R&D Systems, AF4607, 1:100).

Techniques: Translocation Assay, Immunostaining, Staining, Incubation, Control