Ephb4 d1c7n

This was performed as described^{56 (link),57 (link)}. The following primary antibodies were used: EPHB4 (D1C7N) (1: 1000, Cell Signaling #14960), EPHB4 (1:500, Abcam #ab73259), phospho-eiF2α S51 (1:1000, Cell Signaling #9721 S), eiF2α (1:000, Cell Signaling #9722), phospho-IRE1(S724) (1:1000, Abcam #ab48187), c-Myc (Y69) (1:5000, Abcam #ab32072), GLUT3 (1:5000, Abcam #ab191071), Calreticulin (1:400, Abcam #ab2907), HMGB1 (Abcam #ab18256), Actin (1:10000, Cell Signaling #5125), GAPDH (1:5000, Cell Signaling #3683), Phospho-Src Family Tyr416 (1:1000, Cell Signaling #2101), Phospho-p38 MAPK Thr180/Tyr182 (1:1000, Cell Signaling #9211), and Phospho-4EBP1 Thr37/46 (236B4) (1:1000, Cell Signaling #2855). Following incubation with horseradish peroxidase-conjugated goat anti-rabbit/mouse (1:5000, Biorad) or goat anti-rat (1:5000, Santa Cruz) for 1 h, the binding of secondary antibodies were detected with the Supersignal West Femto Maximum Sensitivity Substrate detection system (Pierce) followed by visualization. Quantification analyses were performed by Biorad ChemiDoc Imager and Bio-Rad Image Lab software.

The following materials were commercially obtained: osimertinib (Chemscene, Monmouth Junction, NJ, USA), gefitinib (Wako Pure Chemical Industries, Osaka, Japan), erlotinib (Wako Pure Chemical Industries), and DMSO (Wako Pure Chemical Industries). Antibodies were obtained from the following sources: EGFR [EP38Y] (Abcam, Cambridge, UK), PARP [46D11] (Cell Signaling Technology, Beverly, MA, USA), EphB4 [D1C7N] (Cell Signaling Technology), β-actin [AC-15] (Sigma-Aldrich, Burlington, MO, USA), and Ki-67 [MIB1] (DAKO, Santa Clara, CA, USA).