Fig. 2.

Effects of ABLIM1-specific knockdown on RANKL-induced osteoclast differentiation and activation. shGFP (control vector)- or shAblim1-infected BMMs were cultured with various concentrations of M-CSF (30 ng/ml) and RANKL (100 ng/ml) for 4 d. (A) TRAP of shGFP- or shAblim1-infected osteoclasts was stained. (B) TRAP⁺-multinuclear mature cells (MNCs) with ≥ three nuclei were counted as mature osteoclasts. (C) Total TRAP activity was measured by absorbance at 405 nm. (D) F-actin ring formation was determined by rhodamine-phalloidin staining at 4 d. Pit formation by shGFP- or shAblim1-infected osteoclasts cultured on hydroxyapatite-coated plates was measured at 7 d. The relative resorbed area is presented (right panel). Data represent the mean ± SD and are representative of at least three experiments. *P < 0.05, **P < 0.01 versus control (shGFP). Scale bar = 200 μm.