Figure 6.

Polyamines and DCA were competitively incorporated into rRelish-N. A, rRelish-N was incubated with rTG in the presence of DCA (1 mm) and different concentrations of spermidine or spermine. Treated proteins were separated by SDS-PAGE and detected by UV illumination and CBB staining. B, the density of bands stained with CBB or modified with DCA fluorescence was quantitated using ImageJ software, and the quantity of DCA incorporation was calculated based on CBB staining. Relative intensity of DCA incorporation at 0 mm spermidine or spermine was defined as 1.0. C, rRelish-N was incubated with rTG in the presence of DCA (1 mm) and different concentrations of the spermine. Treated proteins were separated by SDS-PAGE and detected by Western blotting using anti-spermine antibody. D, the bands of spermine-incorporated rRelish-N were quantified by ImageJ software. Relative intensity of spermine-incorporated rRelish-N at 100 mm spermine was defined as 1.0. E, rRelish-N was incubated with rTG in the presence of DCA (1 mm) and different concentrations of spermine. Treated proteins were separated by SDS-PAGE and detected by UV illumination. F, the quantity of DCA incorporation was measured by ImageJ software. Relative intensity of DCA incorporation at 0 mm spermine was defined as 1.0. Data for A–F are representative of three independent experiments.