primary antibodies against stat3  (Affinity Biosciences)

Journal: Frontiers in Endocrinology

Article Title: Chaihu-Wendan Decoction alleviates obesity through PTEN-mediated uncoupling of metabolic signaling and macrophage activation

doi: 10.3389/fendo.2026.1779657

Figure Lengend Snippet: Effect of CHWD on protein expression of STAT3, BCL2, HIF1A, AKT, and mTOR in omental tissues of different groups of mice. Representative Western blots (A) and densitometric analysis of (B) STAT3, (C) HIF1A, (D) AKT, (E) mTOR, and (F) BCL2. Data are expressed as mean ± SD (n = 3). Statistical differences are based on P < 0.05 (*) and P < 0.01 (**) vs. Control group and based on P < 0.05 (#) and P < 0.01 (##) vs. Model group.

Article Snippet: Antibodies and Kits: Primary antibodies against STAT3 (AF6294, 1:1000), HIF1A (AF1009, 1:1000), mTOR (AF6308, 1:1000), p-mTOR (AF3308, 1:1000), BCL2 (AF6139, 1:1000), p-IRS1 (AF3272, 1:1000), p-PI3K (AF3242, 1:1000), and PTEN (AF5447, 1:1000) were purchased from Affinity Biosciences (Cincinnati, OH, USA).

Techniques: Expressing, Western Blot, Control

Journal: Scientific Reports

Article Title: Methylation-induced suppression of BEX1 activates AKT/ERK/STAT3 signaling pathways regulating cell cycle and apoptosis in glioma

doi: 10.1038/s41598-025-14123-8

Figure Lengend Snippet: Bioinformatics Analysis of BEX1 Expression in Glioma Patients. ( A ) Pan-cancer analysis showing significant downregulation of BEX1 in high-grade and low-grade gliomas. ( B , C ) Survival analyses indicating poorer disease-free survival and overall survival in cohorts with low BEX1 expression compared to those with high BEX1 expression. ( D ) Methylation analysis revealing worse survival outcomes in patients with low methylation levels. ( E ) Binary classification of patients based on critical methylation thresholds presented visually. ( F ) Violin plots illustrating the distribution of CpG methylation levels in relation to clinical factors such as age and sex. ( G ) BEX1 mRNA expression levels were analyzed in a cohort of glioma patients ( n = 50), categorized by histopathological grade. A significant downregulation of BEX1 was observed in WHO IV gliomas compared to WHO II ( p < 0.0001, unpaired two-tailed t-test). Data are presented as mean ± SEM. ( H ) Patients with IDH-mutant gliomas were divided into high and low BEX1 expression groups based on the median expression level. Higher BEX1 expression was associated with improved overall survival ( p < 0.01, Log-rank test). ( I ) In MGMT-methylated gliomas, patients with high BEX1 expression also exhibited significantly longer survival than those with low BEX1 expression ( p < 0.05). The number of patients in each group is indicated. ( J ) The network illustrates the potential interactions between BEX1 and associated signaling proteins, including AKT1, MAPK1 (ERK2), and STAT3. Nodes represent proteins and edges represent evidence-based predicted associations. Interaction confidence scores were set to medium (≥ 0.4). The network was visualized using STRING v12.

Article Snippet: Sequential staining was performed using primary antibodies against p-STAT3 (CST, Cat. No. 9145, 1:100), p-ERK1/2 (CST, Cat. No. 4370, 1:200), p-AKT (CST, Cat. No. 4060, 1:100), CDK1 (Abcam, Cat. No. ab32094, 1:100), and BEX1 (Proteintech, Cat. No. 13608-1-AP, 1:100).

Techniques: Expressing, Methylation, CpG Methylation Assay, Two Tailed Test, Mutagenesis

Journal: Scientific Reports

Article Title: Methylation-induced suppression of BEX1 activates AKT/ERK/STAT3 signaling pathways regulating cell cycle and apoptosis in glioma

doi: 10.1038/s41598-025-14123-8

Figure Lengend Snippet: Methylation suppresses BEX1 expression and activates the AKT/ERK/STAT3 signaling pathway to regulate the cell cycle in glioma cells. ( A ) Western blot analysis of p-AKT, p-ERK1/2, and p-STAT3 in U251 and LN229 cells following SAM-induced methylation, with or without BEX1 knockdown or overexpression. Total AKT, ERK1/2, and STAT3 served as internal references. ( B ) Inhibitor validation assays: U251 and LN229 cells were treated with SAM and specific inhibitors of AKT (AKTi), ERK1/2 (ERK1/2i), or STAT3 (STAT3in). The expression levels of phosphorylated proteins and downstream regulators (Cyclin A, CDK1) were detected by Western blot. ( C–D ) qRT-PCR quantification of BEX1, CyclinA, CDK1 ( C ), and CyclinB, CyclinD, CyclinE ( D ) expression in U251 cells across four groups (NC, SAM, SAM + shBEX1, SAM + OE-BEX1). ( E–F ) Corresponding mRNA expression profiles in LN229 cells. Data are presented as mean ± SEM ( n = 3 per group). * P < 0.01, P < 0.05 compared with NC or as indicated (one-way ANOVA with Tukey’s post hoc test).

Article Snippet: Sequential staining was performed using primary antibodies against p-STAT3 (CST, Cat. No. 9145, 1:100), p-ERK1/2 (CST, Cat. No. 4370, 1:200), p-AKT (CST, Cat. No. 4060, 1:100), CDK1 (Abcam, Cat. No. ab32094, 1:100), and BEX1 (Proteintech, Cat. No. 13608-1-AP, 1:100).

Techniques: Methylation, Expressing, Western Blot, Knockdown, Over Expression, Biomarker Discovery, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: Methylation-induced suppression of BEX1 activates AKT/ERK/STAT3 signaling pathways regulating cell cycle and apoptosis in glioma

doi: 10.1038/s41598-025-14123-8

Figure Lengend Snippet: Methylation and BEX1 Influence Glioma Metastasis in Animal Models. ( A ) Subcutaneous tumor formation experiments showing increased tumor burden with methylation and BEX1 knockdown, contrasted with reduced burden upon BEX1 overexpression. ( B ) Relative quantification of tumor burdens. ( C–D ) Ki67 immunohistochemical staining and quantification of proliferating cells in xenograft tissues across indicated groups. ( E ) In vivo imaging of pulmonary metastasis revealing enhanced metastatic capability associated with methylation and BEX1 knockdown, mitigated by STAT3 pathway inhibition. ( F ) Relative quantification of lung metastasis outcomes. ( G ) Representative colony formation assay in U251 and LN229 cells under different conditions (Vehicle, SAM, SAM + sh-BEX1, SAM + sh-STAT3). ( H–I ) Quantification of colony numbers in U251 ( H ) and LN229 ( I ) cells. ( J–K ) CCK-8 assay showing cell proliferation (OD 450 nm) of U251 ( J ) and LN229 ( K ) cells under the same treatments. Data are presented as mean ± SEM. * P < 0.05, ** P < 0.01 (one-way ANOVA with Tukey’s post hoc test).

Article Snippet: Sequential staining was performed using primary antibodies against p-STAT3 (CST, Cat. No. 9145, 1:100), p-ERK1/2 (CST, Cat. No. 4370, 1:200), p-AKT (CST, Cat. No. 4060, 1:100), CDK1 (Abcam, Cat. No. ab32094, 1:100), and BEX1 (Proteintech, Cat. No. 13608-1-AP, 1:100).

Techniques: Methylation, Knockdown, Over Expression, Quantitative Proteomics, Immunohistochemical staining, Staining, In Vivo Imaging, Inhibition, Colony Assay, CCK-8 Assay

Journal: Scientific Reports

Article Title: Methylation-induced suppression of BEX1 activates AKT/ERK/STAT3 signaling pathways regulating cell cycle and apoptosis in glioma

doi: 10.1038/s41598-025-14123-8

Figure Lengend Snippet: Expression Analysis of BEX1 in Glioma Tissue Samples. ( A - F ) Western blot analysis confirming significantly reduced BEX1 protein levels in tumor tissues compared to adjacent non-tumor tissues from 12 glioma patients. ( G-I ) qRT-PCR analysis validating significantly decreased BEX1 mRNA levels in tumor tissues relative to adjacent non-tumor samples across all cases. ( J ) Representative immunofluorescence images of WHO grade II and IV glioma tissues stained for p-AKT, p-ERK, p-STAT3, CDK1, BEX1, and DAPI. ( K ) Quantification of relative fluorescence intensity showing increased p-AKT, p-ERK, p-STAT3, and CDK1, and decreased BEX1 in WHO grade IV gliomas ( n = 3). ( L ) Co-immunoprecipitation in LN229 and U251 cells co-transfected with Flag-BEX1 and His-AKT. BEX1 physically associates with AKT, as shown by reciprocal IP with anti-Flag and anti-His antibodies. Data are shown as mean ± SEM. * P < 0.05, ** P < 0.01; ns: not significant.

Article Snippet: Sequential staining was performed using primary antibodies against p-STAT3 (CST, Cat. No. 9145, 1:100), p-ERK1/2 (CST, Cat. No. 4370, 1:200), p-AKT (CST, Cat. No. 4060, 1:100), CDK1 (Abcam, Cat. No. ab32094, 1:100), and BEX1 (Proteintech, Cat. No. 13608-1-AP, 1:100).

Techniques: Expressing, Western Blot, Quantitative RT-PCR, Immunofluorescence, Staining, Fluorescence, Immunoprecipitation, Transfection