Fig. 6
From: Tumor- and osteoclast-derived NRP2 in prostate cancer bone metastases
Addition of GM-CSF blocks the differentiation and fusion of osteoclasts. Knockout of NRP2 in osteoclastic precursors isolated from NRP2 Fl/Fl; CSF1R-Cre transgenic mice by addition of 4-HydroxyTamoxifen in vitro and differentiated into osteoclasts in the presence of RANKL and M-CSF, LNCaP C4-2B CM and PC3 CM. Under these conditions, one set as control and the other set treated with GM-CSF at 600 pg·mL−1. TRAP staining to compare the osteoclast differentiation in NRP2WT and NRP2KO condition in RANKL+M-CSF at a Day 2. b Day 3. in LNCAP C4-2B CM e Day 2 f day 3. Quantification and comparison of the number of TRAP-positive multinucleated osteoclast per well in NRP2WT and NRP2KO osteoclasts with and without GM-CSF in RANKL and MCSF c Day 2 d Day 3 and in LNCaP C4-2B CM g Day 2 h Day 3. All values reported as mean± SEM from two independent experiments. Statistically significant P value denoted as *** (0.001), **** (0.000 1), ***** (0.000 001). i Flow cytometric analysis of surface markers of mononucleated bone marrow cells differentiated by PC3 CM
