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. 2009 Mar 2;119(4):813–825. doi: 10.1172/JCI36809

Figure 5. The RRI peptide regulates cytoskeleton integrity and cell survival of OCs.

Figure 5

(A) Effect of the RANK peptide on pit formation on dentine slices by mature OCs. OCs were generated on dentine slices and stained with hematoxylin for visualization of pit formation. The area of resorption pits was measured with Image-Pro Plus 4.5 (Media Cybernetics). Data represent mean ± SD. Scale bar: 200 μm. (B) Mature OCs were generated with RANKL and M-CSF, then treated with the RANK inhibitor (WT-1; 5 μM) or control peptide (Mt-1; 5 μM) for 10 hours. Cells were stained for TRAP (upper panels) and F-actin (lower panels). Data represent mean ± SD. n = 6. Scale bar: 100 μm. (C and D) OC survival assays. BMMs were treated with RANKL and M-CSF for 4 days to promote OC formation. On day 4, the cultures were washed with PBS and then treated as indicated. After 10 hours, the cultures were stained for TRAP (C), and a TUNEL assay was performed (D). Results are presented as the mean percentage of apoptotic cells. Data represent mean ± SD. n = 6. Scale bars: 100 μm. (E) Cell lysates from mature OCs were incubated with the caspase-3 (left) or caspase-9 (right) substrate. The release of pNA was measured by absorbance at 405 nm. *P < 0.001. †P < 0.01. Data represent mean ± SD and are representative of at least 3 experiments.