Fig. 2

Effects of Angptl1 on osteoclast formation in mouse bone marrow cells. a Mouse bone marrow cells were pre-cultured with 50 ng/ml of M-CSF for 3 days, and further cultured with 50 ng/ml of RANKL and M-CSF in the presence or absence of recombinant Angptl1 (100 ng/ml) for additional 4 days. The number of TRAP-positive MNCs was counted. Data represent mean ± SEM (n = 6 in each group). **p < 0.01 by Tukey-Kramer test. b Mouse bone marrow cells were pre-cultured with 50 ng/ml of M-CSF for 3 days, and further cultured with 50 ng/ml of RANKL and M-CSF in the presence or absence of recombinant Angptl1 (100 ng/ml) for additional 4 days. Then, total RNA was extracted for qPCR analysis of TRAP, cathepsin K (Ctsk), NFATc1, MMP-9, DC-STAMP or GAPDH. Data are expressed relative to GAPDH mRNA value. Data represent mean ± SEM (n = 6 in each group). Two independent experiments were performed. c Mouse bone marrow cells were pre-cultured with 50 ng/ml of M-CSF for 3 days, and then cultured with or without recombinant Angptl1 (100 ng/ml) for 1 h in the presence of 50 ng/ml of RANKL and M-CSF. Total proteins were extracted from the cells and Western blotting analyses for phosphorylated-p65 (p-p65), p65 and GAPDH were performed. The images represent experiments performed independently 3 times. Cont; Control