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. 2008 Oct 24;113(10):2202–2212. doi: 10.1182/blood-2008-06-162594

Figure 4.

Figure 4

Down-regulation of PIAS3 enhances RANKL-mediated osteoclastogenesis. (A) RAW264.7 cells transfected with 30 nM negative control or PIAS3 siRNA were cultured with 50 ng/mL RANKL for 2 days. Real-time PCR showed that PIAS3 siRNA decreased PIAS3 mRNA by 80%. (B) Analysis of osteoclast differentiation by TRAP staining. RAW264.7 cells were transfected with 30 nM negative control or PIAS3 siRNA and were stimulated by various concentrations of RANKL as indicated. (C) TRAP-positive MNCs having more than 3 and 10 nuclei were separately counted. (D) RAW264.7 cells transfected with negative control or PIAS3 siRNA were cultured with 50 ng/mL RANKL for the indicated times. Total RNA was recovered from cultured cells, and real-time PCR was performed with primers specific for PIAS3, c-Fos, NFATc1, cathepsin K, and TRAP. Error bars in all panels represent SD. #P < .05; ##P < .01; ###P < .001; n.s, not significant.