Fig. 1: DNMDP induces complex formation between purified PDE3A and SLFN12.

a SEC analysis of complex formation performed at 150 mM NaCl. Traces are shown for 10 µM PDE3A^CAT (red), SLFN12 (green), and PDE3A^CAT + SLFN12 in the absence (blue) and presence of 100 µM DNMDP (black). The trace for SLFN12 had to be collected at 500 mM NaCl and is therefore shown as a dashed line. b SE-AUC analysis of PDE3A^CAT and c SLFN12. The solid lines represent the best fit to a monomer–dimer model with the K_d indicated. d Amylose resin pulldown analysis of complex formation. SDS-PAGE gel of protein(s) eluted from amylose resin when 2 µM of His₆-MBP-SLFN12 was incubated alone or with 2 µM PDE3A^CAT ± 10 µM DNMDP or trequinsin at 150 or 500 mM NaCl. e SEC analysis of complex formation performed at 500 mM NaCl. f Effect of DNMDP and trequinsin on complex formation by BLI. Binding of 100 nM SLFN12 to immobilized PDE3A^CAT in the presence of 500 mM NaCl and DMSO (blue), 10 µM DNMDP (black), or 10 µM trequinsin (red). g Quantitative analysis of the effect of DNMDP on complex formation using BLI. Binding to SLFN12 performed in the absence (open circles) and presence (filled circles) of 100 µM DNMDP at 500 mM NaCl. Data were analyzed from four independent experiments and represented as mean values ± s.e.m. Abs. absorbance, Treq. trequinsin.