FIGURE 7.

Influence of AZIN2 deleted forms on polyamine uptake by COS7 cells. A, scheme of the AZIN2 deletions. N-terminal deletions (Δ³⁹N; Δ¹¹³N), C-terminal deletions (Δ⁴⁵C; Δ¹⁶²C), and antizyme-binding site deletion of AZIN2 (Δ^117–140) were obtained by PCR and subcloning into the pcDNA3 plasmid containing the FLAG epitope (see “Experimental Procedures”). B, cells were transfected with the wild type AZIN2 or the different constructs generated. Control cells were transfected with the empty vector. Polyamine uptake assays were carried out using 2 μm ¹⁴C-labeled putrescine for 30 min. Data are shown as mean ± S.E. of triplicate determinations. Protein levels of the different AZIN2 truncated forms were determined by Western blot using anti FLAG antibody. Loading controls were performed using anti Erk2 antibody (data not shown). Ct indicates wild type AZIN2.