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. 2020 May 17;10(15):6638–6660. doi: 10.7150/thno.44793

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Pharmacological activation of Hh signaling by PM restrained osteoclast-specific genes expressions under RANKL stimulation in vitro. qPCR analysis of relative mRNA levels of osteoclast-specific genes, including Nfatc1 (A), c-Fos (B), Ctsk (C), Dcstamp (D), Acp5 (E), Oscar (F), Atp6v0a3 (G), and Atp6v0d2 (H), in BMMs stimulated with RANKL for 6 days in the presence of 0, 0.5, 1, or 2 μM PM. Expression of each target gene was normalized by 18S ribosomal RNA. The relative changes in mRNA level were analyzed by 2^-ΔΔCT method. All values were calculated from three independent biological replicates and presented as mean ± SD. **P<0.01, compared with vehicle-treated group.