Fig. 2.

Liraglutide reduced tumor‐infiltrated NETs. The LLC and Hepa1‐6 cells bearing C57BL/6 mice were treated using PBS or liraglutide, and neutrophils were enriched for in vitro study. Representative images of tumor‐derived NETs from (A) LLC and (C) Hepa1‐6 cells bearing C57BL/6 mice. Blue is nuclear stained by Hoechst 33342. Green is DNA stained by SYTOX Green. Quantification of tumor‐derived NETs from (B) LLC and (D) MC38 cells bearing C57BL/6 mice were conducted using a fluorescence microplate reader (n = 3). The tumor infiltrated neutrophils from (E) LLC and (G) Hepa1‐6 cells bearing C57BL/6 mice were stained with total reactive oxygen species (ROS) assay kit, ROS production was analyzed by flow cytometry, and the green fluorescence of ROS was detected using FITC channel. Mean fluorescence intensity (MFI) of ROS in tumor‐derived neutrophils from (F) LLC and (H) MC38 cells bearing C57BL/6 mice were conducted using a fluorescence microplate reader (n = 3). Nonparametric tests were utilized to assess statistical significance between different treatment groups. *P < 0.05, ***P < 0.001. Error bars represent SD. Scale bar: 400 μm (A, C).