Vincristine vcr

Oxaliplatin (L-OHP) and vincristine (VCR) were purchased from Selleck Chemicals (Houston, TX, USA). Fetal bovine serum (FBS), RPMI-1640 culture medium, penicillin, and streptomycin were purchased from Gibco (Carlsbad, CA, USA). shFOXM1 (target sequence, 5'- CTCTTCTCCCTCAGATATA-3'), shDVL2 (target sequence, 5'-GGAAGAAATTTCAGATGAC-3'), shSnail (target sequence, 5'- GCCTTCAACTGCAAATACT-3'), and shRNA negative control (shNC) were gained from Genechem (Shanghai, China). cDNAs encoding FOXM1 or Snail was respectively cloned into pcDNA3.1 or pcDNA3.1-HA. pcDNA3.1-Flag-DVL2 was derived from pCMV5-3XFlag-DVL2 which was a gift from Jeff Wrana (Addgene plasmid # 24802) [78 (link)]. Lipofectamine 3000 transfection reagent was purchased from Invitrogen (Carlsbad, CA, USA). BCA protein assay kit, Radioimmunoprecipitation (RIPA) lysis buffer, Nuclear and Cytoplasmic Protein Extraction kit, and protein A+G agarose beads were obtained from Beyotime Biotechnology (Nantong, China). Primary antibodies against FOXM1, E-cadherin, N-cadherin, Vimentin, Snail and P-gp were purchased from Cell Signaling Technology (Danvers, MA, USA). Primary antibodies against DVL2, HA, Flag, GAPDH and Lamin B1 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The horseradish peroxidase (HRP)-conjugated secondary antibody was purchased from ZSGB-bio (Peking, China).

Cytarabine (Ara-C) (Selleckchem, Cat # S1648), Methotrexate (MTX) (Selleckchem, Cat # S1210), Vincristine (VCR) (Selleckchem, Cat # S1241), MG132 (Selleckchem, cat # S2619), Caffeine (Sigma-Aldrich, Cat # C0750), and 79-6 (Calbiochem, Cat # 197345) were diluted and stored per manufacturer recommendations. For in vitro experiments drug stocks were diluted in base media and for in vivo experiments stocks were diluted in saline immediately prior to use. In vitro concentrations of Ara-C [1 μM], MTX [50 μM], VCR [25 μM], MG-132 [1-5 μM], Caffeine [2.5-10 mM], and 79-6 [125 μM] were used to approximate clinically relevant doses in ALL or published in vitro concentrations [27 (link), 57 (link)- 63 (link)].