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. 2021 Aug 21;12(2):924–938. doi: 10.1016/j.apsb.2021.08.018

Figure 5.

Figure 5

Transportation of nanoparticles from neutrophils to tumor cells under inflammation. (A) Schematic pattern of neutrophils and the 4T1 coculture model. DiD-labeled neutrophils were incubated with MM-LPs-DiO for 1 h, and then the resulting neutrophils-DiD/MM-LPs-DiO were incubated with tumor cells with PMA for 8 h followed by DAPI staining and microscope observation. (B) Imaging of the coculture model. Yellow and green arrows represent MM-LPs-DiO colocalized with neutrophil-DiD and tumor cells, respectively. Scale bar: 20 μm. Data are representative of three biological replicates. (C) Schematic illustration of 4T1 cells ingesting MM-LPs-DiO released from neutrophils in tumors from mice with or without LPs-R848 treatment. Tumors were collected for IF and flow cytometry experiments. (D) MM-LPs-DiO released from neutrophils and subsequently internalized by the spleen, liver and tumor cells was studied by IF of tissue slices. Yellow and green arrows represent MM-LPs-DiO colocalized with neutrophil-DiD and tumor cells, respectively. The scale bars are 20 μm for the amplification panels and 50 μm for all other panels. Data are representative of three biological replicates. (E) Flow cytometry analysis and (F) corresponding quantitative analysis of the percentage of DiO+ cells in transferred DiD+ neutrophils and CD45- tumor cells (4T1) (n=3). Data are shown as the mean±SD and analyzed by unpaired two-tailed Student's t-test. ∗P<0.05, ∗∗P<0.01.