2 difluoromethylornithine

4-[Aminoiminomethyl]-2,3-dihydro-1H-inden-1-diaminomethylene-hydrazone (SAM486A: also referred to as Sardomozide or CGP 48664) (0.4 µM in H₂O) was provided by Novartis. N1-guanyl-1,7-diamineheptane (GC7) (10 µM in H₂O) was purchased from LGC Biosearch Technologies. As recommended by the manufacturer, GC7 was used together in cell culture with 0.5 mM aminoguanidine to prevent destruction by monoamine oxidase (in H₂O). Deferiprone (DEF) (250 µM in H₂O) was purchased from Calbiochem. Ciclopirox (CPX) olamine (30 µM in H₂O), 2-difluoromethylornithine (DFMO) (200 µM in dimethyl sulfoxide [DMSO]), and N,N1-bis(2,3-butadienyl)-1,4-butanediamine (MDL) (50 µM in DMSO) were purchased from Sigma.
The following antibodies for immunoblots were used (the sources and dilutions shown in parentheses): rabbit anti-VP30 N-terminal region (prepared by GenScript; 1:5,000), rabbit anti-hypusine (Raghavendra Mirmira, Indiana University School of Medicine [35 (link)] and EMD Millipore; 1:1,000), mouse anti-GFP (Roche; 1:1,000), mouse anti-β-actin (Santa Cruz; 1:1,000), mouse anti-VP35 6c5 (Kerafast; 1:1,000), rabbit anti-NP (Integrated Biotechnologies; 1:2,000), rabbit anti-L (Integrated Biotechnologies; 1:1,000); IRDye secondary antibodies: donkey anti-mouse 680 and donkey anti-rabbit 800 (LI-COR Biosciences; 1:10,000).

MTAP− (M−), MTAP+ (M+), and D220A cells were created by stably transfecting either the pTRE2:MTAP, pTRE2:MTAP:D220A, or pTRE2 empty plasmid into HT1080 cells (containing a homozygous MTAP deletion) and pooling 10 individual expressing clones together as was previously described (Tang et al. 2012 (link)). HT1080 cells (Clontech Laboratories, Mountain View, CA) were cultured in Dulbecco’s modified Eagle medium (DMEM) medium supplemented with 2 mM glutamine, 100 μg/mL penicillin, 100 μg/mL streptomycin, 10% fetal bovine serum, and 250 μg/mL G418. Clones were selected using 250 μg/mL hygromycin from a 50 mg/mL stock solution in phosphate-buffered saline (PBS; Sigma-Aldrich, St. Louis, MO). MT-DADMe-ImmA was used at a concentration of 10 μM for all experiments and was obtained from Dr. Vern Schramm (Albert Einstein Medical Center, Bronx, NY). MTA, putrescine, and 2-difluoromethyl-ornithine (DFMO) were obtained from Sigma Aldrich. All media, serum, and antibiotics were obtained from the tissue culture facility at Fox Chase Cancer Center.