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. 2012 Apr;26(4):1484–1492. doi: 10.1096/fj.11-194399

Figure 4.

Figure 4.

Knockdown of Orai1 blunts induction of cytosolic and nuclear NFATc1 during RANKL-mediated osteoclastogenesis. A) Infected RAW cells were cultured with or without RANKL for 2 d, and qRT-PCR analysis of NFATc1 message level was performed. NFATc1 was normalized to GAPDH. Statistical significance was evaluated by an ANOVA followed by Newman-Keuls multiple comparison test. Data are means ± se of triplicate samples from 3 independent experiments with similar results. B) Left panel: Western blot (WB) analysis of cytosolic and nuclear fractionation prepared from infected cells cultured with or without RANKL for 2 d. Actin and lamin B were used as markers of cytosolic and nuclear fractions, respectively. Middle panel: densitometric analysis of cytosolic (middle panel) protein levels. Right panel: densitometric analysis of nuclear NFATc1 protein levels. Data are means ± se of 3 independent experiements with similar results. C) qRT-PCR analysis of the expression of osteoclastic genes known to be a direct transcriptional target of NFATc1 in osteoclastogenesis. Data are means ± se of triplicate samples from 3 independent experiments with similar results. *P < 0.05, **P < 0.01, ***P < 0.001.