Overexpression of Tusc2 in bone marrow-derived macrophage-like cells (BMMs) enhances RANKL-induced NF-κB activation and CaMKIV/CREB signaling. (A) Bone marrow-derived macrophage-like cells (BMMs) were transduced with control or Tusc2 retrovirus. Transduced BMMs were serum-starved for 6 hours and stimulated with RANKL for the indicated times. Whole cell lysates were then harvested from the cultured cells and immunoblotted with the indicated antibodies. (B, C) BMMs were transduced with control or Tusc2 retrovirus. Transduced BMMs were serum-starved for 6 hours and stimulated with RANKL for the indicated durations. Whole cell lysates were then harvested from the cultured cells and immunoblotted with the indicated antibodies. (D) BMMs were transduced with pMX-IRES-EGFP (control) or Tusc2 retrovirus and cultured for 2 days with M-CSF and RANKL. Cytoplasmic fractions and nuclear fractions were harvested from cultured cells and immunoblotted with the indicated antibodies. Antibodies for actin and lamin B1 were used for the normalization of cytoplasmic and nuclear extracts, respectively.