(A) In vitro phosphatase assay. Purified Myc protein was phosphorylated by GSK3β with 32P-γATP. The phosphorylated proteins were incubated with FLAG-Eya1, analyzed on SDS-PAGE, and exposed by phosphorimager.
(B) Eya1 dephosphorylates Myc at T58 but not S62. Purified Myc was incubated with either buffer (−) or FLAG-Eya1 at 30°C for 15 or 30 min. Samples were run in triplicate and immunoblotted with anti-pT58, anti-pS62, or anti-FLAG as indicated.
See also Figure S8.a