Fig. 6.

Gefitinib and PHA665752 co-treatment or SHP2 knockdown impairs U87MG tumor xenograft growth. Mice were subcutaneously injected with control or SHP2-depleted U87MG-M cells. When tumors reached an average size of 50 mm³ (control shRNA only), mice were treated daily either with vehicle or 100 mg/kg gefitinib + 30 mg/kg PHA665752 (G+P) for 7 days (black arrow indicates the start of the treatment). (A) After treatment concluded, pictures were taken and tumors were harvested. (B) Tumor volumes were measured before and throughout treatment. Data are shown as mean ± s.e.m. (control shRNA: vehicle, SHP2 shRNA: no treatment and control shRNA: G + P; n = 12, 26 and 14 tumors, respectively). (C) Tumor lysates were analyzed by western blotting using antibodies against the indicated proteins. Densitometry data are shown as mean ± s.e.m. (n = 3). (D) U87MG-M cells expressing control or SHP2-targeting shRNA were pretreated with DMSO or 40 µM U0126 (control shRNA only) for 24 h prior to hypoxic culture for 24 h. Lysates were analyzed by western blotting using antibodies against the indicated proteins. Densitometry data are shown as mean ± s.e.m. (n = 3); *P<0.05.