Antiphospho aktser473 4060

The EGFR inhibitor Erlotinib, IGF1R inhibitor NVP-AEW541, ERK 1/2 inhibitor PD184352 and mTOR inhibitor Rapamycin (Sirolimus) were all purchased from Selleck Chemicals (California, USA). Drugs were dissolved in DMSO at recommended concentration and stored at −20°C, and diluted in culture medium respectively with <0.1% concentration of DMSO. EGF and IGF1 were purchased from Sino Biological Inc. (Beijing, China), and dissolved in water with final concentration at 100 and 50μg/L, respectively. All primary antibodies used were purchased from Cell Signaling Technology (Boston, USA), which included anti phospho-EGFR^Tyr1068#3777, anti EGFR#2085, antiphospho-IGF1Rβ^Tyr1316#6113, antiIGF1Rβ #9750, antiphspho-MEK1/2^Ser217/221 #9154, anti MEK1/2 #4694, antiphospho-Erk1/2^{Thr202/Tyr204} #8201, antiErk1/2 #9102, antiphospho-Akt^Ser473 #4060, anti Akt#4691, antiphospho-mTOR^Ser2448 #5536 and anti mTOR #2983 antibodies.

Lycium barbarum polysaccharide powder was purchased from Nanjing Manhay Medical Technology (Nanjing, China, No. zhe B2-20090288-37).
DMEM/F12, 0.25% trypsin-ethylenediaminetetraacetic acid, collagenase IV, fetal bovine serum (FBS), and penicillin-streptomycin liquid were purchased from Gibco (BRL, Gaithersburg, MD, USA). 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), dimethyl sulfoxide (DMSO), phosphate buffered saline (PBS), and the bicinchoninic acid (BCA) protein assay kit were purchased from Solarbio Life Sciences (Beijing, China). SP link detection kits (biotin-streptavidin HRP detection systems) and the diaminobenzidine kit were purchased from ZSGB-BIO (Beijing, China). The anti-Ki67 (ab16667) and anti-AR (ab133273) were purchased from Abcam (Cambridge, UK). The anti-occludin (PA5-20755) and anti-zonula occludens-1 (61-7300) were obtained from Thermo Fisher Scientific (Waltham, MA, USA). The anti-Phospho-Akt (Ser473) (#4060) and anti-Akt (#4691) were obtained from Cell Signaling Technology (Boston, MA, USA). The anti-CK-18 antibody (10830-1-AP) and anti-beta-actin antibody (60008-1-lg) were obtained from Proteintech (Chicago, IL, USA).