Phospho c abl

Imatinib was provided by Novartis Pharma AG (Basel, Switzerland). Antibodies for Western blotting, including caspase-9, caspase-3, cleaved caspase-3, PARP, phospho-c-Abl, phospho-Elk-1, phospho-cyclin-dependent kinase 1 (Cdk1) (Thr161), phospho-Cdk1 (Tyr15), phospho-ERK1/2, ERK1/2, phospho-Akt, Akt, phospho-STAT3, STAT3, Bcl-2, and Bcl-xL, were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibodies specific for c-Abl, multidrug resistance 1 (MDR1), α-tubulin, cyclin B1, Cdk1, Mcl-1, Bax, cytochrome c, and β-actin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). An antiphosphospecific MPM2 monoclonal antibody was purchased from Upstate Biotechnology (Lake Placid, NY, USA).

Dasatinib was purchased from Selleckchem, 50 mM and 1 mM stock solutions were made in sterile dimethylsulfoxide. Imatinib was obtained from Novartis (Basel, Switzerland), 2 mM stock solution was prepared in sterile water. fibronectin fragment (120 kDa cell attachment region) was purchased from Chemicon International (CA, U.S.A.). To prepare a fibronectin-coated plate, 50 µl of fibronectin fragment solution (20 µg/ml in sterile water) was added to each well of a Nunc Maxisorp 96-well microtitration plate or of a 16-well E-plate used for real-time cell adhesion monitoring. The plates were subsequently incubated overnight at 10°C. After incubation, the plates were washed three times in PBS and blocked in 1% bovine serum albumin (BSA) in PBS (200 µl/well, 30 min at room temperature). The plate was washed in PBS once again immediately before use.
Antibodies against phospho-SRC (Y417) family (#2101) and phospho-c-ABL (Y245, #2861) were purchased from Cell Signaling. Antibodies against HCK (#610278) and LYN (#610003) were purchased from BD Biosciences, antibody against LCK (Y123, #ab32149) was purchased from Abcam.

Western blotting assay was performed as previously described [15 (link)]. We purchased the following antibodies from Cell Signaling Technology of USA: c-ABL(#2862), Phospho-c/ABL(#2864), HA-Tag(#3724), DYKDDDDK-Tag(#8146), PARP(#9532), Caspase-3(#9662), BID(#2002), Phospho-AKT(#9271), AKT(#9272), Phospho-STAT5(#9351), p73(#14620), Bax(#5023) and p21(#2947S). The HSP90-beta antibody (Genetex, #101448 USA) and β-actin antibody (BOSTER, bs-0061R China) were the same as the antibodies used above with 1:1000 dilution.