Primary human keratinocytes were exposed to either 100nM rapamycin over a timecourse (A), or differing doses of Rapamycin for 24hr (B) and then harvested for Western blot analysis, 20ug per lane. Blots were probed for p-Akt (S473), p-S6 Ribosomal Protein, or beta tubulin as a loading control. HaCaT human keratinocytes were grown to 70% confluence and then treated with rapamycin, PHT-427, or both in serum-free media at the indicated doses. After 24hr the cells were lysed and processed for Western blot analysis, 20ug per lane (C). Blots were probed for p-Akt (S473), p-S6 Ribosomal Protein or beta tubulin as a loading control.