Figure 2

Soluble factors produced by prostate cancer cells increase osteoclast formation from RANKL-primed precursors. A, B) Schematic overviews of experimental setup for osteoclastogenesis assay, from (A) RAW 264.7 and (B) bone marrow cells. RAW 264.7 monocytes were first primed with 50 ng/ml RANKL (RL) for 2 days, and bone marrow cells for 3 days; then the osteoclast precursors (Pre-OC) were cultured untreated (negative control), treated with RANKL (positive control), or supplemented with 10% PC3 or LNCaP CM. C, D) Representative images of osteoclasts formed in different conditions from RAW 264.7 (C), and bone marrow (D) cells. E, F) Average osteoclast numbers formed in different conditions from RAW 264.7 (E) and bone marrow (F) cells. Data are means ± SEM; n = 10 experiments for RAW 264.7 cells, n = 7 for bone marrow cells; *P < 0.05, ***P < 0.001 indicate significance compared to negative control as assessed by Student’s t-test.