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. 2016 Dec 26;8(6):9739–9751. doi: 10.18632/oncotarget.14185

Figure 1. Co-culture of macrophages and prostate cancer cells increases prostate cancer cell migration and invasion and induces CCL2 secretion.

Figure 1

A. U937 differentiation to M2 macrophages is determined using western blot analysis. CCR7 (an M1-type macrophage marker) and CD206 (an M2-type macrophage marker) are assayed using proteins extracted from U937 cells treated with PMA for 24 h and exposed to CM of prostate cancer cells. B. Prostate cancer cells are placed in transwell inserts, and CM of U937 and U937-M cells is added. C. Prostate cancer cells are placed in transwell inserts with Matrigel®-coated membranes in the upper compartment, and U937-M cells are placed in the lower compartments. After 24-h incubation, the cells that had migrated through the membrane are stained. The mean optical density (OD) value is read using a microreader at 595 nm. Data are presented as mean ± SD. D. Chemokine arrays comparing the CM of PC-3 cells in monoculture and the CM of PC-3 cells co-cultured with monocyte cells. E. Prostate cancer cells are co-cultured with U937 cells for 24 h, CM is collected, and CCL2 levels are analyzed using ELISA. Adjustments of brightness, contrast, and size are applied to the whole images of western blot-based analyses without elimination of any information present in the original, including backgrounds. The mean OD value is read using a microreader at 450 nm, and data are presented as mean ± SD. All experiments are performed in triplicate, and the mean values are shown. *p < 0.05, **p < 0.01, ***p < 0.001.