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. 2020 Jun 26;11:3236. doi: 10.1038/s41467-020-17052-4

Fig. 3. Nauclefine binds PDE3A without affecting its phosphodiesterase activity.

Fig. 3

a Pull-down assay of DNMDP-2L binding to Myc-tagged PDE3A (aa 669–1141) WT and to the indicated mutant variants. This is a representative result from three independent experiments. b Myc-tagged PDE3A (aa 669 to 1141) WT and the indicated mutant variants were immunoprecipitated with Myc-beads. The bound PDE3A proteins were detected by western blotting. The bound nauclefine was quantified via LC–MS/MS. The presented result (mean ± SD) is representative of two experiments with three replications in each experiment. Student’s t test (two-tailed, unpaired) was performed. The indicated concentrations of trequinsin and nauclefine (c), or DNMDP and nauclefine (d) were added to the insect-cell purified PDE3A protein in the presence of cAMP for 90 mins. At the end of the assay, the remaining cAMP was measured by LC–MS/MS. This is a representative result (mean ± SD) from two independent experiments with three replicates each time. Student’s t test (two-tailed, unpaired) was performed: *p < 0.05; **p < 0.01; ***p < 0.001; ns not significant.