Cci 779

Nelarabine, ZSTK-474, IPI-145, trametinib, CCI-779, and ABT199 were kindly provided by Selleckchem (Houston, TX, USA). Ara-G, insulin, transferrin, sodium selenite (ITS), propidium iodide (PI), AMD3100, and LY294002 were purchased from Sigma-Aldrich (St. Louis, MO, USA). For western blotting analysis, all the primary antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA) except for the antibodies to CXCR4 and p-CXCR4 (Ser339) which were from Abcam (Cambridge, UK) and antibodies to dGK and dCK which were from Thermo Scientific (Thermo Scientific, Waltham, MA, USA). AlexaFluor^@-conjugated antibodies were from Cell Signaling Technology or BD Biosciences (Franklin Lakes, NJ, USA).

The HK-2 (presented by Prof. Weidong Wang, Zhongshan School of Medicine, Sun Yat-sen University) cells were cultured in DMEM/F12 medium supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin (all from Life Technologies, Carlsbad, CA) and maintained at 37 °C in 5% CO₂ atmosphere. Specifically, HK-2 cells were subjected to starvation in a serum-free medium overnight, then treat with OA (O1383, Sigma-Aldrich) (120 μM) and insulin 2.4 μM for 24 h, and separately subjected to treated with MK-2206 2HCI (S1078, Selleck Chemicals) 1 and 10 μM, CCI-779 (NSC 683864, Selleck) 1 and 10 μM for 24 h. The control group always cultured in a serum-free medium. The experiment was repeated three times. HK-2 cells were stained using SREBP-1 (1:200), BODIPY (1:1000) antibodies and secondary antibodies conjugated with Alexa Fluor. These cells were counterstained with DAPI, and their fluorescent signals were visualized using fluorescence microscope (LSM 800, Zeiss).