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. Author manuscript; available in PMC: 2018 Nov 1.
Published in final edited form as: Amino Acids. 2017 Aug 17;49(11):1793–1804. doi: 10.1007/s00726-017-2477-3

Figure 6. The effects of GC7 on the DHSR1/NADH-Glo (A) and DHSRC (B) assays.

Figure 6

The inhibitory effect of GC7 was tested at 0.3, 1, 3, 10, 30 and 100 μM in the DHS partial reaction and the complete reaction. A, DHSR1 mixtures contained NAD (100 μM), spermidine (20 μM), 0.5 μg of enzyme and indicated concentrations of GC7. After incubation of DHSR1 mixtures at 37 °C for 2 h, NADH-Glo reagent was added and incubated for 30 min at RT before luminescence measurements. B, DHSRC assay was performed in 20 μl, under parallel conditions as DHSR1 except that the eIF5A precursor (3.4 μg), spermidine (3.6 μCi (0.1 nmol) of [1,8-3H]spermidine and 0.3 nmol of unlabeled spermidine) and the enzyme (0.1 μg) were used for each sample. The experiment was repeated with similar results and a representative experiment (in duplicate) is shown.