Fig. 2.

PINK1 inhibits osteoclastogenesis through the Ca²⁺-NFATc1 axis. (A) BMMs isolated from Pink1 WT or Pink1 KO mice were subjected to RT-PCR and western blotting. ***P < 0.001. (B) Pink1 WT or Pink1 KO BMMs were cultured in the presence of RANKL and M-CSF for 3 days and stained for TRAP. The number of TRAP⁺ multinucleated cells with more than 3 or 10 nuclei are presented. MNC, multinucleated cells. Scale bar, 200 μm. (C) BMMs isolated from Pink1 WT and Pink1 KO mice were cultured with the osteoclastogenic medium for 2 days. The mRNA levels of indicated genes were analyzed by RT-PCR. (D) Representative images of dentine slices. The BMMs from Pink1 WT and Pink1 KO were cultured on dentin slices, and the images of dentin slices were obtained by using confocal microscope. *P < 0.05, **P < 0.01. Scale bar, 75 μm. (E) Pink1 WT and Pink1 KO BMMs were cultured with osteoclastogenic medium for 2 days and subjected to Fluo-4/AM staining. Scale bar, 50 μm. (F) Total lysates of Pink1 WT and Pink1 KO BMMs cultured under osteoclastogenic conditions for 2 days were subjected to western blotting. The density of bands relative to α-Tubulin was quantified by ImageJ. *P < 0.05 (G) Pink1 WT and Pink1 KO pOCs were serum-starved and stimulated with RANKL (500 ng/ml) for 15 min. Representative confocal images are presented. Scale bar, 20 μm. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)