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. 2015 Jun 4;35(13):2265–2277. doi: 10.1128/MCB.00062-15

FIG 5.

FIG 5

NOS1AP binds to and modulates the phosphorylation of YAP. (A) Rat brain lysate was precipitated with pan-NOS1AP or Scribble antibodies. Coprecipitated complexes were analyzed by immunoblotting with YAP antibodies (top) and pan-NOS1AP (bottom). (B) HEK293T cell lysate was precipitated with pep-NOS1APc or GST-NOS1APc antibodies. The resulting precipitated complexes were probed with YAP (top) or pep-NOS1APc (bottom) antibodies. (C to F) HEK293T cells were transfected with increasing amounts of a plasmid encoding YFP-NOS1APa (C and D), YFP-NOS1APc (E), or the YFP-PTB domain alone (F). The lysates were analyzed for endogenous Lats1 and YAP and with phospho-Lats1 (detecting S909) and phospho-YAP (detecting S127) antibodies. (G and H) IEC-18 cells transfected with NOS1AP-targeting siRNA (siNOS1AP) (G) or control siRNA (Csi) (H) were lysed at the indicated time points. Equal amounts of cell lysate were prepared for Western blotting and analyzed with the indicated antibodies.