Normal rabbit and mouse igg

Cells were collected and lysed with RIPA buffer plus protease inhibitor cocktail (Roche Applied Science) [2 (link)]. For the chase assay of DR5 protein stability, CHX (100 μg/ml) was introduced for the indicated times. For IP, 10% of the lysate was set aside as the input control [2 (link)]. Cleared lysates were incubated with the indicated antibodies (1 μg) and subsequent Protein A-coupled magnetic Dynabeads (50 μl from a stock solution; Thermo Fisher Scientific). Samples were subjected to standard SDS-PAGE, and the resulting bands were transferred onto polyvinylidene fluoride membrane for visualizing specific proteins [2 (link)].
Primary antibodies used in our study are described as followings: IL13Rα1 (ab79277, Abcam), IL13Rα1 (sc101382, Santa-Cruz Biotech), Trail (27,064, Proteintech), DR5 (ab199357, Abcam), ChoP (15,204, Proteintech), STAT6 (5397, Cell Signaling), phosphor-STAT6 (Tyr641) (5397, Cell Signaling), Bax (50,599, Proteintech), and Bcl (12,789, Proteintech). We used normal rabbit and mouse IgG (Santa Cruz Biotechnology, Inc.) as IP controls. The secondary antibodies used were HRP-coupled anti-mouse IgG and anti-rabbit IgG (GE Healthcare). Antibody binding was detected by enhanced chemiluminescence with hyperfilm ECL or an RGB 600 Imager (GE Healthcare).

Roswell Park Memorial Institute (RPMI) 1640, fetal bovine serum (FBS), Alexa Fluor^TM 488 phalloidin, lipofectamine RNAiMAX, and Halt protease and phosphatase inhibitors were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Purified active MET (the intracellular region of MET, comprising amino acids 974–1390) was obtained from Millipore Sigma (St. Louis, MO, USA). Protein A-Sepharose and glutathione-Sepharose were obtained from GE Healthcare (Chicago, IL, USA). Crizotinib was purchased from Selleck Chemicals (Houston, TX, USA). IQGAP1 siRNA (sc-35700), control siRNA (sc-37007), and normal rabbit and mouse IgG were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Recombinant human Abl1 and Abl2 were obtained from Abcam (Cambridge, UK). Recombinant human HGF was obtained from R&D Systems (Minneapolis, MN, USA). The primary antibodies used in this study are in Table S1. Secondary antibodies were purchased from LI-COR (Lincoln, NE, USA).