Figure 1

Immunoblot characterization of polyclonal antibody IU-88. A , Recombinant human eIF5A was treated in vitro with spermidine, DHS, and DHH as indicated, then subjected to polyacrylamide gel electrophoresis and immunoblots analysis using antibody IU-88 or a pan-anti-eIF5A antibody (a BD monoclonal antibody); B , Recombinant human eIF5A was treated in vitro with spermidine, DHS, and different concentrations of GC7 as indicated, then subjected to polyacrylamide gel electrophoresis and immunoblot analysis using antibody IU-88 or a pan-anti-eIF5A antibody; C , INS-1 β cells were transfected with a plasmid encoding either GFP-eIF5A(K50A) (lane 1) or GFP-eIF5A (lane 2), then cell extracts were subjected to polyacrylamide gel electrophoresis and immunoblots analysis using a pan-anti-eIF5A antibody; D , INS-1 β cells were transfected with a plasmid encoding either GFP-eIF5A(K50A) (lane 1) or GFP-eIF5A (lane 2), then cell extracts were subjected to polyacrylamide gel electrophoresis and immunoblots analysis using antibody IU-88; E , 293T cells were transfected with a plasmid encoding GFP-eIF5A, with or without another plasmid encoding DHS (as indicated), and treated with different concentrations of GC7 as indicated. Cell extracts were then subjected to polyacrylamide gel electrophoresis and subsequent immunoblots analysis using antibody IU-88, a pan-anti-eIF5A antibody, and an anti-actin antibody; F , INS-1 cells were transfected with a plasmid encoding GFP-eIF5A, with or without another plasmid encoding DHS (as indicated), and treated with different concentrations of GC7 as indicated. Cell extracts were then subjected to polyacrylamide gel electrophoresis and subsequent immunoblots analysis using antibody IU-88, a pan-anti-eIF5A antibody, and an anti-actin antibody.