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. 2006 Nov 23;25(24):5840–5851. doi: 10.1038/sj.emboj.7601430

Figure 2.

Figure 2

SPHK1 inhibits RANKL-induced osteoclastogenesis in BMM single culture. (A) BMMs were infected with retroviruses encoding SPHK1 siRNA or luciferase (Luc) siRNA. SPHK1 protein and mRNA levels were evaluated by Western blotting and RT–PCR. (B, C) Infected BMMs were cultured for 4 days in the presence of RANKL (100 ng/ml) and M-CSF (30 ng/ml). Cells were TRAP-stained and the number of TRAP+ MNC was counted. **P<0.005 versus Luc siRNA control. (D) BMMs were infected with retroviruses harboring HA-SPHK1 or the control viruses. The expression of SPHK1 was tested by Western blotting with anti-HA. (E, F). Infected BMMs were induced to differentiate to osteoclasts. The number of TRAP+ MNC was scored. *P<0.01 versus control.