Bms 911543

Detailed description can be found in Supplemental Materials and Methods for: cell line cultures conditions; antibodies and gRNA sequences used; MS instrument settings; in vivo PET imaging Cell lines, SILAC labeling, and reagents Human breast cancer cell lines SKBR3, BT474, BT549, and MDA-MB-231; human non-small cell lung cancer cell lines H292 and HCC827; and HEK-293T were obtained from American Type Culture Collection (ATCC). SUM149 cells were a gift from Prof. W.T.A. van der Graaf (Medical Oncology, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands). Stable isotope labeling of cell lines (SILAC) was done using RPMI or DMEM-high glucose media with normal Arg and Lys (light) or Arg 10 and Lys 8 (heavy) (Silantes). Human MUC1 shedding was measured using a standardized MUC1 human ELISA kit (EHMUC1; Thermo Scientific), mouse MUC1 was measured using a MUC1 mouse ELISA kit (E-EL-M2604; Elabscience). MUC1 has a reported serum half-life of ~7 days 47 . The following inhibitors were used: EGFR inhibitors erlotinib (LC Laboratories; Axon Medchem), gefitinib (Axon Medchem), lapatinib (LC Laboratories), afatinib (Tocris); JAK2 inhibitor BMS-911543 (Selleckchem); PI3K inhibitor BEZ235, AKT inhibitor MK2206, mTORC1/2 inhibitor everolimus, ERK inhibitor FR18024, MEK1 inhibitor AZD6244 (Axon Medchem).