unconjugated anti batf rabbit mab  (Cell Signaling Technology Inc)

Journal: Neoplasia (New York, N.Y.)

Article Title: OSMR coordinates a self-perpetuating circuit linking chemoresistance and neutrophil-driven immunosuppression in gastric cancer

doi: 10.1016/j.neo.2026.101279

Figure Lengend Snippet: Tumor induced PD-L1 expression in neutrophils enhances IL31 secretion of CD8 + T cells. (A) IL31 and OSM mRNA level in CD8 + T cells after co-cultured neutrophils and GC cells. (B) TIM-3, PD-1 and IL31 mRNA level in CD8 + T cells treated with IL2 and Anti-CD3/CD28 antibody. (C) IL31 mRNA and protein level in CD8 + T cells co-cultured with neutrophils treated with BMP5 and PD-L1 inhibitor (Atezolizumab). (D) Schematic of PD-1/PD-L1 signaling pathway in CD8 + T cells. (E) BATF mRNA and protein level in CD8 + T cells treated with IL2 and Anti-CD3/CD28 antibody for 0 day and 14 days. (F) BATF mRNA and protein level in CD8 + T cells after co-cultured neutrophils and GC cells. (G) IL31 mRNA and protein level in CD8 + T cells when BATF was up-regulated or down-regulated. (H) Dual-Luciferase Reporter Assay analysis for BATF-binding IL31 promoter. (I) p-PI3K, PI3K, p-AKT, AKT, p-P27, P27, CCNE2, PD-L1, and PD-L2 expression in MKN28 cells treated with supernatant of CD8 + T cells (J) mIF staining analysis of correlation of OSMR, CD15, CD8, and IL31 in tumor samples from patients in Cohort 2. (**, P < 0.01).

Article Snippet: The membranes were blocked with 5% non-fat milk in TBST for 1 hour at room temperature, followed by an overnight incubation at 4°C with primary antibodies (OSMR (Abcam, #ab315388), CCNE2 (Abcam, #ab40890), β-actin (CST, #4967), PI3K (P85) (CST, #4292), p-PI3K (p-P85) (Abcam, #ab278545), AKT (CST, #9272), p-AKT (CST, #4060), P27 (CST, #2552), p-P27 (Abcam, #ab62364), FLAG (CST, #14793), BMP5 (Proteintech, #13253-1-AP), Ub (CST, #20326), TRIM2 (Proteintech, #20356-1-AP), GATA3 (CST, #5852), p-SMAD1/5 (CST, #9516), SMAD5 (CST, #12534), PD-L1 (CST, #13684), BATF (CST, #8638), IL-31 (abcam, #ab62579)), which were diluted in accordance with the manufacturer's instructions.

Techniques: Expressing, Cell Culture, Luciferase, Reporter Assay, Binding Assay, Staining

Journal: Nature Communications

Article Title: Chromatin remodeling enhances MAP3K8 expression in HAM: a key pathogenesis for therapeutic intervention

doi: 10.1038/s41467-025-64836-7

Figure Lengend Snippet: A Fosl2 and HTLV-1 Tax enrichment (ChIP-seq) in HCT-4 cells and chromatin accessibility (ATAC-seq) in HTLV-1-infected CD4 + T cells in patients with HAM, highlighting the HAM-characteristic OCR. Genomic DNA from HCT-4 cells prior to immunoprecipitation served as input control. ChIP-seq and ATAC-seq peaks were visualized with IGV (data range: ChIP-seq, 0-30; ATAC-seq, 0-100). B Enrichment analysis of transcription factor motifs in ChIP-seq with Fosl2 using the HOMER algorithm. Motifs are ranked by -log10(P). Statistical analysis was performed using a one-sided hypergeometric test. Nominal P values are reported. C Enhancer assays using the MAP3K8 promoter and HAM-characteristic OCR at the MAP3K8 locus. Relative luciferase activity for Fosl2 with partner factors (c-Jun, JunB, BATF, and BATF3) was examined. Data are presented as mean values + SD. Statistical significance was assessed by one-way ANOVA followed by two-sided Tukey’s honestly significant difference (HSD) test for multiple comparisons. Asterisks indicate statistical significance (*** P < 0.001). n = 3, 4, or 5 biologically independent experiments, each performed on separate days using independently transfected HEK293 cells. D Enhancer assays using the MAP3K8 promoter and HAM-characteristic OCR, with Fosl2, c-Jun, and HTLV-1 Tax. Yellow arrows indicate OCR orientation (5’ → 3’ or 3’ → 5’). Data are presented as mean values + SD. Statistical significance was assessed by one-way ANOVA followed by two-sided Tukey’s HSD test for multiple comparisons. Exact P value are as follows: MAP3K8 promoter with Fosl2, c-Jun, and HTLV-1 Tax vs MAP3K8 promoter and HAM-characteristic OCR (5’ → 3’) with Fosl2, c-Jun, and HTLV-1 Tax, P = 0.000010, MAP3K8 promoter with Fosl2, c-Jun, and HTLV-1 Tax vs MAP3K8 promoter and HAM-characteristic OCR (3’ → 5’) with Fosl2, c-Jun, and HTLV-1 Tax, P = 0.000021. Asterisks indicate statistical significance (*** P < 0.001). n = 4 or 7 biologically independent experiments, each performed on separate days using independently transfected HEK293 cells. E Representative histograms depicting chromatin accessibility at the MAP3K8 locus, as determined by ATAC-seq analysis, across various CD4⁺ T cell populations: resting normal CD4⁺ T cells, HAM_D subset, CD4⁺ T cells transiently expressing HTLV-1 Tax for 7 days via a lentiviral system (Tax-cell), and CD4⁺ T cells activated with anti-CD3/CD28 antibodies ( GSM5554060 ). The highlighted region shows HAM-characteristic OCR. ATAC-seq peaks were visualized with IGV (data range: 0-100).

Article Snippet: Anti-c-Jun antibody (9165S, Cell Signaling Technology) Anti-JunB antibody (sc-8051, Santa Cruz Biotechnology) Anti-BATF antibody (8638 T, Cell Signaling Technology) Anti-BATF3 antibody (GTX130125, Gene Tex) Anti-Tax antibody (LT-4, kindly gifted by Dr. Yuetsu Tanaka) Anti-Rabbit IgG (whole molecule)-Peroxidase antibody (A9169, Sigma-Aldrich) Anti-Mouse IgG (whoke molecule)-Peroxidase antibody (A5278, Sima-Aldrich) ECL Prime Western Blotting Detection Reagent (Cytiva, Little Chalfront, England)

Techniques: ChIP-sequencing, Infection, Immunoprecipitation, Control, Luciferase, Activity Assay, Transfection, Expressing