Figure 1

Soluble factors produced by prostate cancer cells do not induce osteoclast formation from naïve RAW 264.7 precursors but increase the cell viability. RAW 264.7 monocytic cells were cultured for 4 days untreated (negative control), treated with 50 ng/ml RANKL (RL, positive control), or supplemented with 10% serum free CM from PC3 or LNCaP, fixed, and stained for TRAP. A) Schematic overview of experimental setup for osteoclastogenesis assay from RAW 264.7 cells. B) Representative images of cultures after 4 days of treatment. C) Average osteoclast numbers formed in different cultures. Data are means ± SEM; n = 10 independent experiments; ***P < 0.001 indicates significance compared to untreated cells as assessed by Student’s t-test. D) RAW 264.7 cells were cultured for 2 days untreated, treated with 50 ng/ml RANKL, or supplemented with 10% PC3 or LNCaP CM. The cells were then incubated with AlamarBlue reagent for 20 h, and cell viability was assessed as the average fluorescence intensity of viable cells subtracted from the readings obtained from the medium. Data are means ± SEM; n = 5 experiments; *P < 0.05 indicate significance compared to negative control as assessed by Student’s t-test.