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Duplication and Diversification of the Spermidine/Spermine N1-acetyltransferase 1 Genes in Zebrafish

Figure 4

Translational regulation inside the ORF of zebrafish family of Ssat1 proteins.

(A) HEK 293T cells were transiently transfected with the plasmid encoding myc-tagged full-length human SSAT1, zebrafish Ssat1a, Ssat1b, or Ssat1c. After incubation for 12 h, transfected cells were treated with 10 µM DENSPM, 20 µM MG132, or vehicle for 24 h. (B) HEK 293T cells were transiently transfected with the plasmid encoding myc-tagged zebrafish Ssat1 chimeric enzymes (detail in Materials and Methods). After incubation for 12 h, transfected cells were treated with vehicle (lane 1 and 3) or 10 µM DENSPM (lane 2 and 4) for 24 h. Cell lysates (5 µg total protein in each lane) were prepared and the protein content of Ssat1 and β-actin in each sample was detected by western blotting with anti-myc and anti-β-actin antibody.

Figure 4

doi: https://doi.org/10.1371/journal.pone.0054017.g004