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