Fig. 6.

Western blotting demonstrates dephosphorylation of Nth1 S83 upon inhibition of PKA, PKA+Ypk1 and Ypk1. Strains wt^as, pka^as, pka^as+ypk1^as and ypk1^as harboring endogenously C-terminally 3xFLAG tagged Nth1, the corresponding isogenic wt strain with 3xFLAG-tagged nth1-S20A,S21A,T58A,S60A,S83A (wt^as nth1–5A-3xFLAG) and untagged isogenic wt strain (wt^as) were treated with DMSO or 500 nm 1NM-PP1 for 15 min. Eluates from immuno-precipitates with an anti-FLAG antibody were probed with antibodies against S83p and against the FLAG-tag.