Cell-based interaction analysis of MOB1A mutants.
A, Interaction profiles of FLAG-MOB1A and the indicated MOB1A N-terminal extension mutants, affinity-purified from stable HEK293 cell lines after treatment with okadaic acid (OA) or DMSO vehicle controls. Statistical analysis of the interaction confidence was performed using SAINTexpress (49) and visualization was through prohits-viz.lunenfeld.ca (see Experimental Procedures). Interactions are listed for the upstream Hippo kinase module, LATS and NDR kinases, DOCK6-8 and PP6 phosphatase modules. See inset legend for details. All interactions are listed in supplemental Table S4 and the underlying protein identifications in supplemental Table S5. Also see ProHits-web.lunenfeld.ca (project: “phospho-dependent Hippo interactions”). B, Control Western blot analysis of FLAG-MOB1A and endogenous PPP6C, LATS1 and LATS2 expression levels.