Female athymic Balb/c nude mice (Taconic Farms), 4–5 weeks old, were housed in a pathogen-free temperature controlled isolation room, with a diet consisting of autoclaved rodent chow and autoclaved water given ad libitum. NCI-H1299 cells (1
*10^7) were injected into the right flank of each mouse by subcutaneous injection. Palpable tumors were observed in approximately 90% of the mice after 1 week. Polyethylene glycol (PEG, 100 Al) or PD176252 (10 or 1 ug in 100 ul of PEG 400) were injected daily by gavage. In anxiety model, PD176252 was administered to rats intraperitoneally at doses 3.75-7.5 mg/kg or intracerebloventicularly, 100-200 ng/rat.

Growth studies in vitro were conducted using the MTT colorimetic assays. NCI-H1299 cells (104/well) were placed in SIT medium and various concentrations of PD176252 added. After 4 days, MTT (Sigma) was added. After 4 h, 150 Al of dimethylsulfoxide was added. After 16 h, the optical density at 570 nm was determined. The IC50 for PD176252 was 5 and 7 AM using NCI-H1299 and H345 cells, respectively.