Fig. 4.

Effects of ABLIM1-specific knockdown on RANKL-mediated osteoclast mobility. (A) Cell migration of shGFP- or shAblim1-infected osteoclasts was measured as described in the Materials and Methods section. Data are expressed as the representative mean ± SD of the number of migrating cells from at least three independent experiments. *P < 0.05 versus control (shGFP). (B) To detect PI3K and AKT activation, shGFP- or shAblim1-infected BMMs cultured with M-CSF (30 ng/ml) overnight were stimulated by RANKL (100 ng/ml). To analyze Rac1 expression, BMMs infected with shGFP-or shAblim1 were cultured under treatment of M-CSF (30 ng/ml) and RANKL (100 ng/ml) for 4 d. Cell lysate (30 μg) was subjected to SDS-PAGE and analyzed by immunoblotting with their respective antibodies. The fold change was normalized to non-phosphorylated proteins (PI3K or AKT) or actin, respectively. Data are representative of three independent experiments.