Fig. 1

c-Kit phosphorylates PHB at tyrosine 259. a Different amounts of active c-Kit kinase domain (0, 20, 40, 80, 160 ng) reacted with 1 μg PHB-His in an in vitro kinase assay. The luminescence was read using a Biotek microplate reader. Data represent the mean ± SD of three independent experiments. ***, P < 0.001; **, P < 0.01; *, P < 0.05, t test. b Activity of c-Kit kinase, phospho-PHB^Y259-His and PHB-His from the in vitro kinase assay was analyzed by western blotting. For preabsorption control, anti-phospho-PHB^Y259 antibody was pre-incubated with an excess amount of the phosphopeptide spanning phospho-tyrosine 259 of PHB (IAYQLSRSRNIT (pY)LPAGQSVLLQ; 10 μg peptide/1 μg antibody) for 2 h at room temperature and then used for western blotting in the rightmost lane. Blots are representative of three independent experiments