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. Author manuscript; available in PMC: 2011 Feb 25.

Published in final edited form as: J Cell Physiol. 2009 Aug;220(2):332–340. doi: 10.1002/jcp.21767

Fig. 5 — RAW264.7 cells were seeded into the 48-well cell culture plates and induced with 50ng/ml RANKL and 5ng/ml M-CSF. Three variables were examined- [HCO₃⁻] (0, 12, and 24 mM), 2-hydroxyestradiol (5 µM) and 8-Br-cAMP (0.1 mM) A. Cells were cultured for 7 days, fixed and stained for TRAP (pink). Images show representative cells collected from one of four independent experiments. B: Effects of HCO₃⁻ on osteoclast size in the background of vehicle (open), 5 µM 2-hydroxyestradiol (light gray), 0.1 mM 8-bromo-cAMP (black), 5 µM 2-hydroxyestradiol & 0.1 mM 8-bromo-cAMP (dark gray). Four images from each well were randomly selected from each of four independent experiments. The area of each TRAP(+)-multinucleated osteoclast was determined using ImageJ software. Bars and error bars represent means ± SEM. * Significant difference compared to 0 mM HCO₃⁻ treated cells (p<0.05).

Fig. 5 — RAW264.7 cells were seeded into the 48-well cell culture plates and induced with 50ng/ml RANKL and 5ng/ml M-CSF. Three variables were examined- [HCO₃⁻] (0, 12, and 24 mM), 2-hydroxyestradiol (5 µM) and 8-Br-cAMP (0.1 mM) A. Cells were cultured for 7 days, fixed and stained for TRAP (pink). Images show representative cells collected from one of four independent experiments. B: Effects of HCO₃⁻ on osteoclast size in the background of vehicle (open), 5 µM 2-hydroxyestradiol (light gray), 0.1 mM 8-bromo-cAMP (black), 5 µM 2-hydroxyestradiol & 0.1 mM 8-bromo-cAMP (dark gray). Four images from each well were randomly selected from each of four independent experiments. The area of each TRAP(+)-multinucleated osteoclast was determined using ImageJ software. Bars and error bars represent means ± SEM. * Significant difference compared to 0 mM HCO₃⁻ treated cells (p<0.05).