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. 2009 Apr 24;284(17):11531–11545. doi: 10.1074/jbc.M808324200

FIGURE 8.

FIGURE 8.

Comparison of cellular PTP-oc protein levels in marrow-derived osteoclasts (A), relative size of TRACP+, multinucleated osteoclasts (B), responsiveness of marrow osteoclast precursors to permissive doses of sRANKL in formation of TRACP+, multinucleated osteoclasts (C), and in vitro bone resorption activity of osteoclasts (D) of young adult male F2 transgenic mice with osteoclasts of male WT littermates. A, Western blot of the PTP-oc protein in two marrow-derived osteoclast cultures of an 8-week-old male transgenic mouse and that in two osteoclast cultures of an age-matched male WT littermates. The same blot was blotted against actin for protein loading control. B, microphotograph of a representative culture of marrow-derived osteoclasts of a 8-week-old transgenic mouse (right) and of an age-matched WT littermate (left) are shown. Scale bar, 20 μm. C summarizes the number of TRACP+, multinucleated osteoclasts formed from marrow-derived osteoclast precursors (at 1 × 104 cells/well in 96-well plates) of 7-week-old male transgenic mice and those of age-matched male WT littermates after 7 days of treatment with 1, 5, or 10 ng/ml sRANKL and 50 ng/ml m-CSF. Results are shown as mean ± S.E. (n = 6). Statistical significance was analyzed with two-factor analysis of variance. D summarizes the in vitro bone resorption activity of the derived osteoclasts from eight male 10-week-old PTP-oc transgenic and eight age-matched male WT littermates on the resorption pit formation assay. Briefly, the same numbers of marrow-derived osteoclast precursors were plated on 5-mm dentine slices in a 48-well plate and were treated with RANKL and m-CSF for 7 days. The cells were then removed by sonication, and the resorption pits were stained with acid hematoxylin. The size of more than 50 resorption pits per slice was measured and divided by the number of pits to get pit area per pit (osteoclasts). The top shows a representative field of resorption pits for each test group. Scale bar, 100 μm. The bottom shows the average pit area per pit (osteoclast) formed (mean ± S.E., n = 8 for each group).