T egfr

Antibodies to the following were used at 1:1000 dilution unless specified: p-EGFR (Y1068), p42/44 pMAPK(T202/Y204), p42/44 MAPK(L34F12, 1:2000), Akt (9272), p-Akt (Ser473, D9E), cleaved PARP (Asp214), PARP (9542), Estrogen Receptor alpha (D8H8) cleaved caspase-3 (Asp 175, 5A1E, 9664), caspase-3 (9662), cleaved caspase-8 (Asp 374, E6H8S,), caspase-8 (1C12, 9746) (Cell Signaling Inc., Beverly, MA, USA) SEMA3C (N-20, 1:2000); Plexin B1 (H300, 1:500), t-EGFR (C-2, 1:1000) (Santa Cruz Biotechnology, Santa Cruz, CA), Actin (A2066, 1:5000), Vinculin (1:5000) (Roche Diagnostics, Laval, PQ, Canada), LICOR secondary antibodies- anti Rabbit IR680 and IR800, anti mouse IR680 and IR800 (Rockland, Gilbertsville, PA, USA) and anti Goat680 (Pierce Biotechnology, Rockford, IL, USA). Antibodies were used according to manufacturer’s protocols. The following chemicals were used: Tamoxifen (68392-35-8; Sigma-Aldrich, Oakville, ON, Canada), Fulvestrant (I4409-25MG; MiliporeSigma, Oakville, ON, Canada), Estradiol (E2758; Sigma-Aldrich, Oakville, ON, Canada), Charcoal Strip Serum (12676029; Gibco, Grand Island, NY, USA), RNase A (12091021; Invitrogen, Carlsbad, CA, USA), Propidium Iodide (P3566; Invitrogen, Carlsbad, CA, USA) and Insulin-transferrin-selenium (41400045; Gibco, Grand Island, NY, USA).

Pra-B (purity > 98%) was purchased from ChemFace (Hubei, China). The origin stock solution of Pra-B is 100 mM in DMSO solvent. The final DMSO concentrations ranged from 0.01% for 10 μM Pra-B to 0.05% for 50 μM Pra-B. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Antibodies for CTSC, CTSV, t-ERK, p-JNK, t-JNK, p-p38, t-p38, p-MEK, t-MEK, p-EGFR, t-EGFR, and β-actin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The antibody for p-ERK was purchased from Cell Signaling Technology (Danvers, MA, USA).