Rabbit anti hypusine

Western blotting was performed as described (58 (link)). The following primary antibodies were used: mouse anti-V5 (1:5,000; Thermo Fisher Scientific, Waltham, MA, USA), mouse anti-tubulin (1:2,000; Solarbio, Beijing, China), rabbit Anti-GAPDH (1:2,000; Solarbio), and rabbit anti-Hypusine (EMD Millipore; 1:1,000) antibodies, rabbit anti-eIF5A (1:1,000; ABclonal), rabbit anti-DHPS (1:200; abcam), rabbit anti-DOHH (1:500; Sigma-Aldrich). Secondary antibodies used were Goat Anti-Mouse horseradish peroxidase-conjugated secondary antibody (1:2,000; Beyotime, Shanghai, China) and Goat Anti-Rabbit horseradish peroxidase-conjugated secondary antibody (1:5,000). Anti-His (M1001020, Solarbio), Anti-pAKT473 (GB13012-3, Servicebio), Anti-pAKT308 (ab66134, Abcam), Anti-ATPaseβ (GL Binchem synthesis), Anti-Inx1(GL Binchem synthesis), Anti-Inx2 (GL Binchem synthesis), Anti-Inx3(GL Binchem synthesis), Anti-Inx4 (GL Binchem synthesis), Cleaved-caspase (WL02117, Wanleibio), Anti-GAPDH (M1000110, Solarbio), Anti-β Tubulin (AF1216, Beyotime), Goat Anti-Mouse (A0216, Beyotime), Goat Anti-Rabbit (A0208, Beyotime). Proteins were semi-quantified via densitometry using ImageJ (National Institutes of Health, Bethesda, MD, USA).

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).