Western Blotting
GIST
Anlotinib with 5 concentrations, 0, 10, 50, 200, and 500 nmol/L, was added to the 2 GIST cell lines for 4 h when the confluence of the cells were 90%. The whole cell lysates were prepared using a standard protocol. The protein lysate concentrations were measured using a Quick Start Bradford 1 × Dye Reagent (Bio-Rad; #5000205). Electrophoresis and western blotting were performed using standard techniques. The hybridization signals were detected by chemiluminescence (Immobilon Western, Millipore Corporation, MA) and captured using an Amersham Imager 600 imagers (GE Healthcare; #29083461).