Fig. 4

Osteoclast‐specific knockout of PDK1 suppressed RANKL-stimulated osteoclastogenesis, bone resorption function, and podosome belt formation in vitro. A BMM viability detected by CCK-8 after 48 h of M-CSF stimulation. B Images of mature osteoclasts after TRAP staining; scale bar = 2000 μm. C Quantification of osteoclasts in PDK1 cKO and WT mice. D Fluorescence images of the effects of PDK1 on the podosome actin belt; scale bar = 1000 μm. E Count of osteoclasts with podosome actin belts. F Quantification of the average number of nuclei per osteoclast. G Images showing hydroxyapatite resorption; osteoclasts were seeded on hydroxyapatite-coated bone absorption plates and stimulated with RANKL. H Quantification of resorbed hydroxyapatite. N = 3, all data are expressed as the mean ± SD. *P < 0.05, **P < 0.01, and ***P < 0.001. BMMs, bone marrow-derived macrophage cells; TRAP, tartrate-resistant acid phosphatase