FIGURE 8.

Analysis of the role of C/EBPα silencing in OC survival. A, C/EBPα silencing attenuates OC survival. MBM cells were first treated with M-CSF (10 ng/ml) and RANKL (10 ng/ml) for 2 days (d) before infection with a lentivirus encoding scramble shRNA control (Scr-sh) or C/EBPα shRNA construct 1 or 5 (C/EBPα-sh 1 or 5) for 1 day in the presence M-CSF (10 ng/ml) plus RANKL (10 ng/ml). The cultures were then continued to complete a 4-, 6-, and 8-day culture. The cultures were stained for TRAP activity. Quantification is shown in the bottom panel from three independent experiments. Scale bars = 200 μm. B, C/EBPα silencing attenuates RANKL-induced Akt activation in MBM cells. MBM cells expressing Scr-sh or C/EBPα-sh 1 or 5 were treated with RANKL (10 ng/ml) as indicated. The activation of the Akt pathway was assessed by Western blotting as phosphorylation of Akt (pAkt) using total Akt (Akt) as a loading control in three independent experiments. The numbers in parentheses show concentration in nanograms per milliliter. Error bars show mean ± S.D. *, p < 0.05.