Cp 724714

HER2 inhibitor CP-724714 was from Selleckchem (Houston, TX, USA). Normal human IgG were purchased from Sigma-Aldrich (St. Louis, MO, USA). Trastuzumab was from Roche (Basel, Switzerland). Mouse monoclonal antibodies to human EGFR (1005), HER2 (A-2) (sc-393712), human EGFR (1005), Akt1/2 (H-136), Erk1/2 (k-23), pT202/Y204 Erk1/2 were from Santa Cruz Biotechnology Inc. (Dallas, TX, USA). Rabbit monoclonal antibodies to phospho-Akt (Ser473) and phosphor(Thr308), and rabbit monoclonal antibody to HER2/ErbB2 (29D8) were purchased from Cell Signaling Technology (Danvers, MA, USA). Rabbit polyclonal antibodies to phospho-HER2 Y-1005, Y-1112, Y-1127, Y-1139, Y-1196, and Y-1248 were from FroggaBio (Toronto, ON, Canada). Secondary antibodies used for Western analysis, including Anti-rabbit and anti-mouse RDye^® 800 CW and RDye^® 650, were from LI-COR biotechnology Inc. (Lincoln, NE, USA). Invitrogen™ Cholera Toxin Subunit B (Recombinant) and Alexa Fluor™ 488 Conjugate were purchased from Fisher Scientific (Ottawa, ON, Canada). All other chemicals were purchased from Sigma-Aldrich (Oakville ON, Canada).

Cell culture media, phosphate buffered saline, and fetal bovine serum were obtained from Invitrogen (Gibco, Karlsruhe, Germany). Antibodies against Akt, phospho-Akt, p44/42 MAPK, phospho-p44/42 MAPK, actin, and anti-mouse IgG Alexa Fluor^® 647 (#4410S) were purchased from Cell Signaling (Danvers, MA, USA). Anti-HER3 purified antibody (clone 1B4C3) was purchased from BioLegend^®. Secondary antibodies were from Sigma–Aldrich (Steinheim, Germany). Protran Nitrocellulose Transfer membranes were purchased from Whatman (Dassel, Germany). The enhanced chemiluminescence systems (Super Signal West Femto Maximum Sensitivity Substrate and SuperSignal West Pico Chemiluminescent Substrate) were from Thermo-Scientific (Bonn, Germany). The WST-1 kit was from Roche Applied Science (Mannheim, Germany). The PCK inhibitor bisindolylmaleimide II (BIM II), the MET inhibitor PF04217903, and the HER1 inhibitor AG1478 were from Tocris (Wiesbaden, Germany). The HER2 inhibitor CP724714 was purchased from Selleckchem (Munich, Germany). Heregulin and 12-O-Tetradecanoylphorbol 13-acetate (PKC activator, PMA) were obtained from Sigma–Aldrich (Steinheim, Germany). The human phospho-MAPK Array Kit was from R&D (Minneapolis, MN, USA). All other chemicals used were purchased from Carl Roth (Karlsruhe, Germany) unless indicated otherwise.

HER2 kinase inhibitor CP-724714 was purchased from Selleckchem (Houston, TX, USA). Pertuzumab (Perjeta^®) and trastuzumab (Hercepton^®) were purchased from Hoffmann-La Roche (Basel, Switzerland). Mouse monoclonal anti-human HER2 (9G6) (sc-08), anti-human HER2 (A-2) (sc-393712), anti-human EGFR (A-10) (sc-373746), anti-human HER3 (RTJ.2) (sc-415), and rabbit polyclonal anti-human pY1248 HER2 (sc-12352-R) antibodies were purchased from Santa Cruz Biotechnology Inc. (Dallas, TX, USA). Rabbit polyclonal anti-human pY1005, pY1112, pY1127, pY1139, and pY1196 HER2 were purchased from FroggaBio (Toronto, ON, Canada) and mouse monoclonal anti-human pY1221/1222 HER2 (6B12) (2243) were from Cell Signaling Technology (Danvers, MA, USA). Anti-rabbit and anti-mouse RDye^® 800CW and RDye^® 650 secondary antibodies were purchased from LI-COR biotechnology Inc. (Lincoln, NE, USA). Isotype control human IgG and all other chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA).