Multiscript rt

Antibodies against Bcr, p-Akt/Akt, PI3K p110β, Stat5, Erk1/2, p-Erk1/2,Bcl-xL, PARP and PathScan^® Bcr/Abl Activity Assay (Phospho-c-Abl, Phospho-Stat5 and Phospho-CrkL Multiplex Western Detection Cocktail) were from Cell Signaling Technology (Andover, MA, USA). Antibodies against GAPDH, β-actin and c-Abl were from Santa Cruz Biotechnology (Santa Cruz Biotechnology, CA, USA), Antibodies against FLAG or HA were from Sigma-Aldrich (St Louis, MO, USA). Secondary antibodies were chosen according to the primary antibodies origin. Proteasome inhibitor MG132 was from Calbiochem (Billerica, MA, USA). Imatinib mesylate was from Department of Hematology, Tangdu hospital, the Fourth Military Medical University. SYBR Premix Ex Taq II and Multiscript RT were purchased from TaKaRa Biotechnology (Dalian) Co., Ltd (Dalian, China).

Human cervical cancer HeLa, liver cancer HepG2, pancreatic cancer PANC-1 and human embryonic kidney cell HEK-293T cell lines were maintained in DMEM (Life Technologies) supplemented with 10% fetal bovine serum. All these cell lines were cultured in a 37°C incubator with 5% CO2 humidified air. Antibodies against IGF-1Rα, IGF-1Rβ, IRβ,β-Actin and α-tubulin were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA), FLAG and IgG were from Sigma-Aldrich (St Louis, MO, USA), and Glut4, Akt, p-Akt(S473), ERK and p-ERK from Cell Signaling (Andover, MA, USA). Antibody against HA was purchased from NeoMarkers (Fremont, CA, USA). Antibody against N-cadherin was from BD Biosciences. IGF-1 and insulin were obtained from Sigma-Aldrich, and the proteasome inhibitor MG132 from Calbiochem (Billerica, MA, USA). SYBR Green universal master mix and Multiscript RT were purchased from TaKaRa Biotechnology (Dalian) Co., Ltd (Dalian, China). Cy3- and FITC-conjugated secondary antibodies and 4’,6-diamidino-2-phenylindole (DAPI) were purchased from Boster Biology Company (Wuhan, China). 2-[N-(7-Nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy-D-glucose (2-NBDG), a fluorescence labeled deoxyglucose analog, was obtained from Cayman Chemical Company (Ann Arbor, MI, USA).

Microglia were labelled using a rabbit primary antibody targeting ionized calcium binding adapter molecule 1 (IBA‐1) (019‐19741) from Wako (Richmond, VA, USA). Antibodies against basigin (Rabbit polyclonal antibody used for Western blot, ab64616; Rat monoclonal antibody used for immunofluorescence double labelling, ab34016), PECAM (ab9498), IGF‐1 (ab9572), IGF‐1 receptor (ab16890), VEGF (ab51745) and VEGFR‐2 (ab5473) were purchased from Abcam (Cambridge, MA, USA). HIF‐1α (MAB 5382) was obtained from Millipore (Billerica, MA, USA). Antibodies against AKT (9272), P‐AKT (4060), ERK (4695), P‐ERK (4370) and the MEK1/2 inhibitors PD98059, PI3K inhibitor LY294002 were from Cell Signaling (Andover, MA, USA). Antibodies against β‐actin were from Santa Cruz (Santa Cruz, CA, USA). Recombinant human IGF‐1 (100‐11) was from ProteinTech (Rochy Hill, NJ, USA). Secondary antibodies including goat anti‐rabbit conjugated to AlexaFluor 594/CY3 or AlexaFluor 488/FITC were purchased from Beijing ComWin (Beijing, China). SYBR Premix Ex Taq II and Multiscript RT were purchased from TaKaRa (Dalian, China).