Fig. 7: Phosphorylation of native FRQ by CK1a triggers its dissociation from FRH.

A Whole cell lysate (WCL) of Neurospora expressing 2 × FLAG-tagged FRH prepared from cultures containing hypophosphorylated FRQ (grown 3 h in light after 10 h darkness) was incubated with recombinant CK1a and ATP, or left untreated. B Lysates from (A) were subjected to FLAG immunoprecipitation. Total (T), supernatant (S), and immunoprecipitate (IP) were analyzed by Western blot with antibodies against FRQ (upper panels and 2 × FLAG-FRH (Lower panels). Results shown are a representative of three independent experiments with reproducible results.