Fig. 2

Overexpression of GPR84 directly inhibits osteoclast formation. A Representative image of flow cytometry (left) and quantification (right) of BrdU incorporation in BMMs treated by 6-OAU (200 nM) or vehicle in the presence of M-CSF (50 ng/mL) (n = 4). B BMMs isolated from bone marrow were treated by 6-OAU (200 nM) or vehicle in the presence of M-CSF (50 ng/mL) and the relative expression of cell cycle biomarkers was determined by qRT-PCR (n = 3). C, D Representative TRAP stain images of BMMs treated with CM from MC-38, 6-OAU or transfected with PCMV6-GPR84 (C) and quantification of osteoclast number (D) (n = 5). Bar represents 50 μm. E BMMs were isolated from bone marrow treated with CM from MC-38, 6-OAU or transfected with PCMV6-GPR84. The relative OCSTAMP level was determined by qRT-PCR (n = 3). F BMMs were isolated from bone marrow treated with CM from MC-38, 6-OAU or transfected with PCMV6-GPR84. The relative Ctsk level was determined by qRT-PCR (n = 3). Significant differences are indicated as *p < 0.05, **p < 0.01 or ***p < 0.001