anti phospho stat4 tyr693  (Cell Signaling Technology Inc)

Journal: bioRxiv

Article Title: The impact of patient biology on racial disparities in breast cancer outcome

doi: 10.1101/2025.11.05.685148

Figure Lengend Snippet: (A) Linear regression plot depicting the relationship between STAT4 expression and patient poverty burden in the VA dataset. (B) STAT4 expression relative to patient poverty burden in the CBCS validation cohort, where the horizontal dotted line indicates average STAT4 expression. (C) tSNE plots from the Single Cell Portal for STAT4 expression depicting highest expression levels in immune T-cell subsets. (D) Representative immunohistochemistry images for phospho-STAT4 protein expression in patient tumor sections from the VA-dataset across two separate patient tumors from high- and low-poverty burden with accompanying quantification represented as stacked columns. (E-F) Kaplan Meier survival curves comparing patient tumors with high and low STAT4 RNA expression for women with estrogen receptor positive/human epidermal growth factor receptor 2 negative breast cancer from Metabric (E, left) and TCGA (E, right & F), with (F) or without race stratification (E). (G) Working model schematic highlighting the interplay between patient-intrinsic and -extrinsic factors in the modulation of tumor biology that contributes to poor patient outcomes in Black breast cancer patients. Scale bar is 290um (D). Linear regression coefficients were used to determine p-values using a T-test (A). Fisher’s exact test was used for determining the p-values in (B and D). Survival analysis was performed using the Mantel-Haenszel method to determine hazard ratios (HR) and a log-rank Mantel-Cox test was used to determine p-values (E-F). Abbreviations: TMM (Trimmed Mean of M values), TPM (Transcripts per million), VA-cohort (Veterans Affairs cohort), CBCS-cohort (Carolina Breast Cancer Study cohort), CD8 (cluster of differentiation 8), CD4 (cluster of differentiation 4), Treg (Regulatory T cells), POV (poverty burden), HR (hazard ratio), n (number).

Article Snippet: Antibodies used were STAT4 (Cell Signaling Technology, catalog no. #4134), and Histone H3 (Cell Signaling Technology, catalog no. #9701).

Techniques: Expressing, Biomarker Discovery, Immunohistochemistry, RNA Expression

Journal: Experimental Hematology & Oncology

Article Title: Improving CAR-T cell function through a targeted cytokine delivery system utilizing car target-modified extracellular vesicles

doi: 10.1186/s40164-025-00701-z

Figure Lengend Snippet: IL-12 EVs enhance the function of CAR-T cells. ( A ) CAR-T cells were treated with control EVs or IL-12 EVs for 30 min, and the phosphorylation of STAT4 was analyzed by western blotting. Experiments are representative of 3 independent repeats with similar results. (B-G) CAR-T cells (cell density: 1*10 6 /mL) were mixed with 5 × 10 4 Raji cells at an effector-to-target ratio of 1:1 and treated with PBS, control EVs, rhIL-12, and IL-12 EVs repectively. Protein concentration of EVs is 167 µg/mL, and rhIL-12 concentration is 667 pg/mL. ( B , C ) Cytokine secretion by CAR-T cells was detected by ELISA after 24 h of coculture ( n = 3 donors). ( D ) The proportion of CAR-T cells was analyzed by flow cytometry after 72 h of coculture ( n = 3 donors). ( E ) CD107a expression in CD8 + CAR-T cells was detected by flow cytometry ( n = 3 donors). ( F ) Raji cell death was determined using PI (BD Pharmingen) and analyzed by using flow cytometry after 24 h ( n = 5 donors). ( G ) Subsets were detected via flow cytometry in CAR-T cells after 7 days of treatment ( n = 3 donors). Data are presented as mean ± SEM. Statistical analysis was performed using one-way or two-way ANOVA. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001

Article Snippet: Total STAT4 and phosphorylated STAT4 in T cells were detected using an anti-Stat4 (C46B10) antibody (CST, 2653 S, 1:1000) and an anti-phospho-Stat4 (Tyr693) antibody (CST, 5267 S, 1:1000).

Techniques: Control, Phospho-proteomics, Western Blot, Protein Concentration, Concentration Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Expressing