Journal: Mediators of Inflammation

Article Title: Aspirin Disrupts the Crosstalk of Angiogenic and Inflammatory Cytokines between 4T1 Breast Cancer Cells and Macrophages

doi: 10.1155/2018/6380643

Figure Lengend Snippet: Effect of aspirin on M1 and M2 macrophage subtypes following LPS stimulation and coculture with 4T1 cells. Macrophages were incubated in the presence or absence of aspirin for 72 h and cultured in either fresh medium as a control, the presence of LPS for the last 24 h of the incubation, or cocultured with 4T1 cells for 72 h. (a) Histogram plots, (b) fluorescent intensity plots, and (c) quantitative data were presented. The immunofluorescent intensity of CD11c (M1) and CD206 (M2) on macrophages was analyzed using a NC-3000. Data are shown as mean ± SEM. Statistical analysis was performed using one-way ANOVA and LSD post hoc tests. The comparisons between different culture mediums were done by t -tests. Statistically significant differences are indicated as ∗ p < 0.05, # p < 0.01, and † p < 0.001 versus vehicle control.

Article Snippet: Macrophages were incubated in the presence or absence of aspirin for 72 h and cultured in either control medium, the presence of LPS for the last 24 h of the incubation, or cocultured with 4T1 cells for 72 h. To assess surface marker expression, RAW 264.7 and 4T1 cells were collected after 72 h of coculturing and stained by incubating with fluorescein FITC anti-mouse CD11c and Alexa Fluor 647 anti-mouse CD206 monoclonal antibodies (Sony Biotechnology Inc.) at 4°C in the dark for 30 min. After washing, viable cells were stained with Hoechst 33342 (ChemoMetec, Allerød, Denmark) and subjected to FlexiCyte fluorescence-activated cell sorting analysis.

Techniques: Incubation, Cell Culture, Control