Fig. 5
Afatinib and TMZ combination reduces tumor burden in vivo. a, b U87EGFRvIII luciferase transfected cells (2 × 104 in 2 μl of PBS) were intracranially injected into 4–6 week’s old mice using a stereotactic frame. After 5 days, mice were randomized into 4 groups and treated with vehicle (n = 8), TMZ (25 mg/kg BW) (n = 7), afatinib (10 mg/kg BW) (n = 8) or combination (n = 5) for 5 days a week by oral gavage. Animals treated with TMZ alone or in combination with afatinib survived longer and were sacrificed after 30 days. The tumor volume (total photon count) was measured using IVIS imaging on days 0, 7, 9, 11, 20 and 30. ($, # P ≤ 0.03); $ significant compared to TMZ; # significant compared to vehicle control (c) Kaplan Meier survival curve analysis showing effect of control, afatinib, TMZ or combination on OS of EGFRvIII orthograft mice. d, e Confocal microscopy showing expression of (d) pEGFR (Tyr-1068) (red staining), Oct3/4 (green staining) and (e) pcMET (Tyr-1234/1235) (red) and Nanog (green) images in U87EGFRvIII tumor xenografts