Figure 2. Disruption of Hedgehog signaling inhibited ex vivo osteoclastogenesis in a cell-autonomous manner.
A) Smofl/fl bone marrow derived macrophages were lentivirally infected with Cre-recombinase or GFP control to delete Smoothened, differentiated into OCs and stained with TRAP. B) Quantification of above OC area per total area on day 6. C) TRAP staining of OC differentiated in the presence of vehicle or LDE225 for 6 days. D) Treatment with 1μM LDE225 decreased Gli1, Nfatc1, Itgb3, and Ctsk transcripts by qRT-PCR on days 3 and 5 of OC differentiation. Data normalized to gene expression of day 1 vehicle treated macrophages. E) Day 3 preOC treated with 1μg/ml recombinant murine Shh for 24 hours had increased levels of Gli1, Nfatc1, and Itgb3 by qRT-PCR.