Modulation of Tumor-Associated Macrophages (TAM) Phenotype by Platelet-Activating Factor (PAF) Receptor

Figure 6

TAMs from PAFR KO tumors show the M1-like phenotype. CD45⁺ cells were purified from tumor suspensions from animals by positive selection using CD45-coated magnetic beads (Miltenyi Biotec) and stimulated with 10 ng/mL of LPS. After 72 hours of incubation at 37°C in 10% SFB RPMI, the culture supernatants were harvested for nitric oxide (nitrite) production detection by Griess reaction (a) and IL-10 detection by ELISA (c). The cellular extracts were used for determination of arginase activity (b) normalized by the total protein concentration measured by BCA kit. indicates for WT compared with PAFR KO tumors. Each experimental group contained 4 animals. pM are peritoneal macrophages treated in exactly the same way as tumor infiltrating cells, used as control for NO detection and arginase activity assays.

(a)

(b)

(c)