Bez235 s1009

The recombinant humanised monoclonal HER2 antibody trastuzumab (a concentration of 15 µg/mL was selected as indicated elsewhere [24 (link)]) (Herceptin, Genentech, San Francisco, CA, United States) was supplied by the pharmacy of our hospital; BEZ235 (S1009), everolimus (S1120) and TAK-228 (S2811) were obtained from Selleckchem (Selleckchem Spain, Madrid, Spain).

Antibodies against ERα (sc-543), and EGFR (sc-03) were from Santa Cruz Biotechnology (Heidelberg, Germany); antibodies against phosphorylated Akt (9271S), mitogen activated protein kinase 42–44 (MAPK) and phosphorylated MAPK (9101 and 137 F5), and phosphorylated EGFR (4407) were from Cell Signalling Technologies (Leiden, The Netherlands); antibody for Akt was a kind gift from P. Coffer (UMC, Utrecht, The Netherlands). For analyzing phosphorylated proteins the Western-Star immunodetection kit (Tropix kit) from Applied Biossytems (Foster City, CA, USA) was used. TAM, fulvestrant, E2, EGF, and the protein dye sulforhodamin B (SRB) were from Sigma Aldrich (St Louis, MO, USA). Mitogen-activated kinase kinase (MEK) inhibitor U0126 (V-112A) was from Promega (Leiden, The Netherlands); Phosphoinositide 3-kinase (PI3K) inhibitor BEZ235 (S1009) was from Selleck (Houston, TX, USA).