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. 2017 Nov 14;7:15575. doi: 10.1038/s41598-017-15586-0

Figure 2.

Figure 2

FAM19A5 blocks RANKL-induced osteoclastogenesis. (A,B) Mouse BMDMs were stimulated with FAM19A5 (2 μM) or LPS in the presence of M-CSF (30 ng/ml) and RANKL (100 ng/ml) for 4 days. All cells were stained with TRAP staining solution. TRAP+ 3 < MNCs (B, left) and TRAP+ 5 < MNCs (B, right) were counted. (C) Mouse BMDMs were stimulated with various concentrations of FAM19A5 (0, 0.01, 0.1, 1, and 2 μM) in the presence of M-CSF (30 ng/ml) and RANKL (100 ng/ml) for 4 days. (D) Mouse BMDMs were pre-incubated with polymyxin B (10 μM) prior to FAM19A5 (2 μM) and LPS (1 μg/ml) treatment in the presence of M-CSF (30 ng/ml) and RANKL (100 ng/ml) for 4 days. (E) Mouse BMDMs were stimulated with 2 μM FAM19A5 (left) or various concentrations of FAM19A5 (0, 0.01, 0.1, 1, and 2 μM) (right) in the presence of M-CSF (30 ng/ml) and RANKL (100 ng/ml) for 4 days. After adding CCK-8 assay solution, the OD was measured at 450 nm. Data in these panels are representative of three independent experiments (A). Data are presented as means ± SE (n = 3). Data are representative of at least three independent experiments (B–E). ns: not significant, *p < 0.05; **p < 0.01.