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 BMP5 activates the SMAD1/5 pathway in neutrophils and promotes PD-L1 transcription. (A) Correlation between neutrophil score and BMP5 receptors in TCGA database. (B) Schematic of BMP5 and its receptors signaling pathway. (C) p-SMAD1/5 and SMAD protein levels in neutrophils. (D) PD-L1 mRNA and protein level in neutrophils treated with BMP5 and SMAD1/5 inhibitor. (E) PD-L1 mRNA and protein level in neutrophils co-cultured with MKN28 cells or treated with SMAD1/5 inhibitor. (F) Dual-Luciferase Reporter Assay analysis for SMAD5-binding PD-L1 promoter. (G) mIF staining and correlation of BMP5,CD15, PD-L1, and CD8 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: Cell Culture, Luciferase, Reporter Assay, Binding Assay, Staining