Journal: MedComm

Article Title: An Engineered Nano‐Bridging Strategy Remodels the Immune Microenvironment of Esophageal Squamous Cell Carcinoma via STING Pathway Activation

doi: 10.1002/mco2.70828

Figure Lengend Snippet: HA1@diABZI–HMGB1 activates the STING signaling pathway in BMDMs and enhances proinflammatory cytokine production. (A) Western blot analysis was conducted to examine STING, TBK1, IRF‐3, and their phosphorylated forms in BMDMs following various treatments. (B) A quantitative assessment of the phosphorylation levels of proteins within the STING signaling pathway was performed. (C) A schematic representation of the experimental design is provided. Primary BMDMs were isolated from murine bone marrow and differentiated using macrophage colony‐stimulating factor (M‐CSF) at a concentration of 20 ng/mL over a period of 7 days. The mature BMDMs were subsequently cocultured with mEC25 tumor cells and exposed to different treatments for 24 h. Culture supernatants were collected and subjected to ELISA to quantify the levels of IFN‐β, IL‐6, TNF‐α, and CXCL10. (D–G) ELISA was utilized to quantify the levels of IFN‐β, IL‐6, TNF‐α, and CXCL10 in the coculture supernatants following the various treatments. All data expressed as mean ± SD ( n = 3). Statistical significance was calculated through one‐way ANOVA using the Tukey's posttest. * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Article Snippet: BMDMs were isolated from 6‐ to 8‐week‐old female C57BL/6 mice and cultured in complete DMEM medium supplemented with M‐CSF at a concentration of 20 ng/mL (Sangon Biotech) at 37°C in an atmosphere containing 5% (v/v) CO 2 .

Techniques: Western Blot, Phospho-proteomics, Isolation, Concentration Assay, Enzyme-linked Immunosorbent Assay