Fig. 3. Recombinant RON13 is active in vitro.

a Scheme of recombinant RON13 kinase (rRON13k) and RON13 dead kinase (rRON13dk), in which kinase activity is abrogated by introducing the mutation D595A. b Coomassie-stained gel of purified rRON13k and RON13dk produced in insect cells. c Radioactive RON13 kinase activity assays based on rRON13k autophosphorylation. Radiograph is shown below the graph. Densitometry data (mean ± SD; n = 3 biologically independent experiments) were normalized to rRON13k in presence of Mn²⁺. Statistical significance was determined using a two-tailed paired t test. STS staurosporine. d ATP hydrolysis during kinase reactions, as measured by TLC analysis (mean ± SD; n = 3 biologically independent experiments). Autoradiograph showing both non-hydrolyzed ATP (ATP) and inorganic phosphate (Pi) is shown below the graph. P values were determined by a two-tailed paired t test. Source data are provided as a Source data file.