CCI-779's effects on p70S6kinase phosphorylation and cell-cycle regulatory proteins in vivo. After 5 days of vehicle alone or CCI-779 treatment (20 mg/kg) in 8226- or OPM-2-challenged mice, tumor nodules were harvested, and extracted protein was immunoblotted for expression of total p70S6kinase (p70), phosphorylated p70, total STAT-3, or phosphorylated STAT-3. This experiment was repeated 2 additional times with identical results. (B) Effects of CCI-779 on expression of cyclin D1, p27, and c-myc. OPM-2- or 8226-challenged mice were treated with vehicle alone or varying doses of CCI-779 (shown above blots as mg/kg) for 5 days, after which tumor nodules were harvested, and extracted protein was immunoblotted for expression of p27, c-myc, cyclin D1, and actin. This experiment was repeated once with identical results.