Raw246.7 macrophages

Caco-2 cells and RAW246.7 macrophages, purchased from BeNa Culture Collection (Beijing, China), were cultured in Dulbecco's modified Eagle's medium (Gibco, Grand Island, NY) containing 10% fetal bovine serum (Clark, Australia) and were maintained at 37 °C in a humidified chamber of 5% CO₂.
The mice were intraperitoneally injected with 4 mL of 3% thioglycollate broth. After three days, the mice were sacrificed and then each mouse was injected with 4 mL RPMI 1640 into the peritoneal cavity. The peritoneal lavage fluid was collected and centrifuged at 300g for 5 min. The supernatant was removed and the cells were re-suspended in RPMI 1640 medium supplemented with 10% fetal bovine serum and maintained at 37 °C with 5% CO₂. After 2 h, non-adherent cells were discarded. The remaining adherent cells were cultured overnight until they were used for the experiments.

Human prostate cancer cell line PC3 and RAW246.7 macrophages were purchased from BeNa Culture Collection (Beijing, China). PC3 cells and RAW246.7 macrophages were cultured in PRMI 1640 medium (Invitrogen, Gaithersburg, MD, USA) containing 10% fetal bovine serum (Invitrogen, USA) under the condition of 37°C and 5% CO₂. IL-4 (10 ng/mL) was used to stimulate RAW246.7 macrophages to obtain anti-inflammatory macrophages (M2),28 (link) while control macrophages (M0) were cultured in medium alone. Pro-inflammatory macrophages (M1) were induced by treating RAW246.7 macrophages with 50 ng/mL interferon (IFN)-γ and 10 ng/mL lipopolysaccharide (LPS).29 (link)