primary antibodies against p stat3  (Cusabio)

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