Journal: Immunity, Inflammation and Disease

Article Title: Interleukin‐35 Ameliorates Obesity‐Associated Impairment of Endothelium‐Dependent Vasodilation via the STAT3‐MAPK/ERK‐eNOS Signaling Axis

doi: 10.1002/iid3.70564

Figure Lengend Snippet: IL‐35 activates the STAT3‐MAPK/ERK‐eNOS signaling axis to promote eNOS phosphorylation and NO production in endothelial cells. (A) Western blot bands of p‐STAT3, p‐eNOS (Ser1177), p‐cRAF, p‐MEK, p‐ERK, and corresponding total proteins in HUVECs. (B–I) Densitometric analysis of p‐STAT3, STAT3, p‐eNOS (Ser1177), eNOS, p‐cRAF, p‐MEK, and p‐ERK normalized to total proteins or β‐tubulin, n = 3. (J–K) NO fluorescence intensity in HUVECs (J, scale bar = 50 μM) with semi‐quantification (K, n = 3). Data are mean ± SD. ns, not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Primary antibodies for immunofluorescence and Western blotting: EBI3 (12371‐1‐AP), IL‐12A (85754‐2‐RR), platelet endothelial cell adhesion molecule‐1 (PECAM‐1/CD31) (11265‐1‐AP), eNOS (27120‐1‐AP), and β‐tubulin (66240‐1‐Ig) from Proteintech (USA); p‐eNOS (Ser1177, #9571), p‐cRAF (#9427), p‐MEK1/2 (#9154), and p‐STAT3 (#9145) from Cell Signaling Technology (USA); cRAF (A19638), MEK1/2 (A4868), p‐ERK1/2 (AP0974), ERK1/2 (A4782), and STAT3 (A1192) from ABclonal Technology (China).

Techniques: Phospho-proteomics, Western Blot, Analysis, Fluorescence

Journal: Frontiers in Neurology

Article Title: Ferroptosis-associated candidate genes in aneurysmal subarachnoid hemorrhage: evidence from human peripheral blood and mouse brain

doi: 10.3389/fneur.2026.1823039

Figure Lengend Snippet: Identification and functional enrichment of ferroptosis-related differentially expressed genes in the GSE36791 peripheral-blood dataset. (A) Heatmap of all differentially expressed genes in control and aSAH samples. (B) Heatmap of the 35 ferroptosis-related differentially expressed genes. (C) Venn diagrams showing the overlap between all genes detected in GSE36791 and human ferroptosis-related genes obtained from FerrDb, and the overlap between GSE36791 differentially expressed genes and FerrDb genes, identifying 35 ferroptosis-related differentially expressed genes. (D) Spearman correlation heatmap of the 35 ferroptosis-related differentially expressed genes. (E) Gene Ontology enrichment analysis of these genes. (F) Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis of these genes. DEGs, differentially expressed genes; Ferr-DEGs, ferroptosis-related differentially expressed genes; GO, Gene Ontology; KEGG, Kyoto Encyclopedia of Genes and Genomes.

Article Snippet: Sections were then incubated overnight at 4 °C with the following primary antibodies: anti-PGD (1:100, Cat. A0563, Abclonal, Wuhan, China), anti-STAT3 (1:100, Cat. A11216, Abclonal), anti-MAPK3/ERK1 (1:100, Cat. A19561, Abclonal), and anti-G6PD (1:100, Cat. A27296, Abclonal).

Techniques: Functional Assay, Control, Analysis

Journal: Frontiers in Neurology

Article Title: Ferroptosis-associated candidate genes in aneurysmal subarachnoid hemorrhage: evidence from human peripheral blood and mouse brain

doi: 10.3389/fneur.2026.1823039

Figure Lengend Snippet: Network-based identification of ferroptosis-associated candidate hub genes in aSAH. (A) PPI network constructed from ferroptosis-related DEGs. (B) Top-ranked PPI hub genes identified by cytoHubba. (C–E) MCODE clusters identified from the PPI network, with scores of 4.286, 4.000, and 3.000, respectively. (F) Intersection between WGCNA module genes and PPI-derived hub genes. (G) Overlap between the four selected hub genes and DCI-associated genes. PPI, protein–protein interaction; DCI, delayed cerebral ischemia.

Article Snippet: Sections were then incubated overnight at 4 °C with the following primary antibodies: anti-PGD (1:100, Cat. A0563, Abclonal, Wuhan, China), anti-STAT3 (1:100, Cat. A11216, Abclonal), anti-MAPK3/ERK1 (1:100, Cat. A19561, Abclonal), and anti-G6PD (1:100, Cat. A27296, Abclonal).

Techniques: Construct, Derivative Assay

Journal: Chemistry & Biodiversity

Article Title: Network Pharmacology and In Vivo Evaluation of Modified Huanglian Wendan Decoction in Polycystic Ovary Syndrome With Insulin Resistance: Association With AGE–RAGE‐Related Signaling

doi: 10.1002/cbdv.71737

Figure Lengend Snippet: Ovarian AGE–RAGE/STAT3/AKT1‐related protein expression and phosphorylation. (A) Representative Western blot bands for AGE, RAGE, STAT3, AKT1, phospho‐STAT3 (Ser727), phospho‐AKT (Ser473), and GAPDH in the Control, Model, HLWDD‐L, HLWDD‐M, HLWDD‐H, and Metformin groups. (B) Relative densitometric analysis of AGE/GAPDH, RAGE/GAPDH, STAT3/GAPDH, AKT1/GAPDH, p‐STAT3/STAT3, and p‐AKT/AKT1. ** p < 0.01 vs. Control; ## p < 0.01 vs. Model. Densitometric comparisons were based on independent biological replicates and analyzed as described in Methods.

Article Snippet: Primary antibodies included AGE (A16422, ABclonal; 1:2000), RAGE (66833‐1‐Ig, Proteintech; 1:2000), STAT3 (60199‐1‐Ig, Proteintech; 1:1000), AKT1 (10176‐2‐AP, Proteintech; 1:2000), phospho‐STAT3 (Ser727; AP0715, ABclonal; 1:2000), phospho‐AKT (Ser473; 80455‐1‐RR, Proteintech; 1:1000), and GAPDH (60004‐1‐Ig, Proteintech; 1:20000).

Techniques: Expressing, Phospho-proteomics, Western Blot, Control, Analysis

Journal: Frontiers in Physiology

Article Title: Kaempferol ameliorates gestational diabetes mellitus by regulating STAT3-associated AIM2 inflammasome activation and ferroptotic injury in trophoblasts

doi: 10.3389/fphys.2026.1857741

Figure Lengend Snippet: KPF ameliorates placental pathological changes and oxidative stress in GDM rats. (A) HE staining was performed to evaluate pathological alterations in placental tissues. Magnification: 100×; scale bar: 200 μm. (B) PAS staining was performed to assess glycogen distribution and deposition in placental tissues. Magnification: 100×; scale bar: 200 μm. (C, D) DHE staining was performed to evaluate ROS accumulation in placental tissues. Magnification: 200×; scale bar: 20 μm. (E–I) Western blot analysis was performed to determine the protein expression levels of AIM2, p-STAT3, GPX4, and SLC7A11 in placental tissues. N = 3, * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Following blocking, the membranes were incubated overnight at 4°C with primary antibodies, including AIM2 (1:1000, catalog number ab93015, Abcam), Glutathione Peroxidase 4 (GPX4, 1:1000, 52455, Cell Signaling Technology, CST), solute carrier family 7 member 11 (SLC7A11, 1:1000, 23276, CST), phospho-STAT3 (p-STAT3, 1:1000, 9145, CST), STAT3 (1:1000, 12640, CST), pro-IL-1β (1:1000, 12703, CST), IL-1β (1:1000, ab283818, Abcam), cleaved caspase-1 (1:1000, 4199, CST), pro-caspase-1 (1:1000, 24232, CST), STAT1 (1:1000, 9172T, CST), phospho-STAT1 (p-STAT1, 1:1000, 9177S, CST), IRF1 (1:1000, 9172T, CST) and β-actin (1:1000, 4967S, CST).

Techniques: H&E Stain, Staining, Western Blot, Analysis, Expressing

Journal: Frontiers in Physiology

Article Title: Kaempferol ameliorates gestational diabetes mellitus by regulating STAT3-associated AIM2 inflammasome activation and ferroptotic injury in trophoblasts

doi: 10.3389/fphys.2026.1857741

Figure Lengend Snippet: KPF modulates trophoblast inflammatory responses and ferroptosis-related alterations through STAT3-associated signaling regulation. (A) Molecular docking analysis showing the potential interaction between KPF and STAT3 protein. (B, C) RT-qPCR analysis of STAT3 mRNA expression and transfection efficiency in HTR-8/SVneo cells. (D) CCK-8 assay was used to evaluate cell viability. (E, F) ELISA quantification of inflammatory cytokines IL-18 and TNF-α in cell supernatants. (G–L) Western blot analysis of AIM2, IL-1β, pro-IL-1β, GPX4, SLC7A11, cleaved caspase-1, and pro-caspase-1 protein expression in cells subjected to various treatments. (M–P) Western blot analysis of STAT3-associated changes in STAT3, GPX4, and SLC7A11 protein expression in cells subjected to various treatments. N = 3, not significant, * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Following blocking, the membranes were incubated overnight at 4°C with primary antibodies, including AIM2 (1:1000, catalog number ab93015, Abcam), Glutathione Peroxidase 4 (GPX4, 1:1000, 52455, Cell Signaling Technology, CST), solute carrier family 7 member 11 (SLC7A11, 1:1000, 23276, CST), phospho-STAT3 (p-STAT3, 1:1000, 9145, CST), STAT3 (1:1000, 12640, CST), pro-IL-1β (1:1000, 12703, CST), IL-1β (1:1000, ab283818, Abcam), cleaved caspase-1 (1:1000, 4199, CST), pro-caspase-1 (1:1000, 24232, CST), STAT1 (1:1000, 9172T, CST), phospho-STAT1 (p-STAT1, 1:1000, 9177S, CST), IRF1 (1:1000, 9172T, CST) and β-actin (1:1000, 4967S, CST).

Techniques: Docking Assay, Analysis, Quantitative RT-PCR, Expressing, Transfection, CCK-8 Assay, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Frontiers in Physiology

Article Title: Kaempferol ameliorates gestational diabetes mellitus by regulating STAT3-associated AIM2 inflammasome activation and ferroptotic injury in trophoblasts

doi: 10.3389/fphys.2026.1857741

Figure Lengend Snippet: STAT3-associated changes in STAT1/IRF1 signaling and AIM2 expression in HG-stimulated trophoblast cells. (A, B) RT-qPCR was performed to assess STAT3 knockdown efficiency and AIM2 mRNA expression changes in HTR-8/SVneo cells subjected to different treatments. (C–F) Western blot analysis was performed to evaluate the expression levels of p-STAT1, STAT1, IRF1, and AIM2 proteins under different experimental conditions. N = 3, not significant, ** P <0.01, *** P <0.001.

Article Snippet: Following blocking, the membranes were incubated overnight at 4°C with primary antibodies, including AIM2 (1:1000, catalog number ab93015, Abcam), Glutathione Peroxidase 4 (GPX4, 1:1000, 52455, Cell Signaling Technology, CST), solute carrier family 7 member 11 (SLC7A11, 1:1000, 23276, CST), phospho-STAT3 (p-STAT3, 1:1000, 9145, CST), STAT3 (1:1000, 12640, CST), pro-IL-1β (1:1000, 12703, CST), IL-1β (1:1000, ab283818, Abcam), cleaved caspase-1 (1:1000, 4199, CST), pro-caspase-1 (1:1000, 24232, CST), STAT1 (1:1000, 9172T, CST), phospho-STAT1 (p-STAT1, 1:1000, 9177S, CST), IRF1 (1:1000, 9172T, CST) and β-actin (1:1000, 4967S, CST).

Techniques: Expressing, Quantitative RT-PCR, Knockdown, Western Blot, Analysis