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. 2012 Jan;32(2):490–500. doi: 10.1128/MCB.06361-11

Fig 5.

Fig 5

Conditioned medium from Akt1-deficient osteoblasts reduces osteoclast formation. (A) Experimental scheme to assess osteoclast differentiation in mouse bone marrow macrophages incubated in a combination of 50% osteoblast conditioned medium (CM, collected at day 7 from control or shAkt1 osteoblasts) and 50% growth medium (GM). (B) Example of TRAP staining to assess formation of multinucleated osteoclasts on day 7. (C) Percentage of TRAP-positive cells expressed as area per microscopic field (×200 magnification, mean ± S.D. of 6 fields, ∗, P < 0.01 versus Con CM). (D) Quantitative assessment of the percentage of multinucleated TRAP-positive osteoclasts on day 7 per microscopic field (×200 magnification; mean ± SD of results of six fields; ∗, P < 0.01; ∗∗, P < 0.001 versus Con CM). Inset depicts representative examples (white arrowheads indicate osteoclasts with >4 nuclei). (E) Expression of mRNAs for cathepsin K, the calcitonin receptor (Rec), and S17 by RT-PCR on day 7.