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. Author manuscript; available in PMC: 2015 Sep 1.
Published in final edited form as: Calcif Tissue Int. 2014 Jun 17;95(3):209–217. doi: 10.1007/s00223-014-9883-2

Fig. 3.

Fig. 3

Knocking down of Tspan-10 expression attenuates osteoclast formation. (A) BMMs were transduced with recombinant lenti-viruses expressing a control shRNA targeting fire fly luciferase (LUC-sh) or two Tspan-10-targeting shRNAs (Tsp10-sh1 and Tsp10-sh2), respectively. After selecting with 6 μg/ml puromycin for 3 days, the cells were cultured with M-CSF alone (BMM) or with M-CSF plus RANKL for 5 days (mOC). Total RNAs were isolated from three independent cultures of each group. Quantitative real-time RT-PCR analysis of the expression of Tspan-10 was performed. * p < 0.05, ** p < 0.01 vs LUC-sh by Student's t-test. (B) Lenti-viruses transduced BMMs were cultured with M-CSF and RANKL for 5 days. The cells were fixed and stained for TRAP. The scale bar = 20 μm. (C) The mRNA expression of osteoclast marker gene, TRAP (encoded by Acp5), was measured by quantitative real-time PCR using TaqMan assay primers from Life Technologies., n = 3, ** p <0.01 vs LUC-sh by Student's t-test. (D) The protein expression of Cathepsin K was detected by western blots. Tubulin served as a loading control.