Analysis of C. elegans DAF-16 phosphorylation in different genetic backgrounds.

C. elegans DAF-16/FOXO phosphorylation in different genetic backgrounds was analyzed by mass spectrometry of affinity purified HA-tagged DAF-16. Genetic backgrounds studied include par-1 hypomorphic mutants, PAR-1 knockdown using an auxin-inducible degron (AID), daf-2(e1370) and akt-1(ok525) mutants. PAR-1 is the C. elegans ortholog of MARK kinases; DAF-2 is the sole C. elegans insulin/IGF receptor, and AKT-1 is one of two AKT kinases in the nematode.