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. 2013 May 3;32(11):1543–1555. doi: 10.1038/emboj.2013.102

Figure 2.

Figure 2

Requirement of cAMP/PKA signalling for induction of YAP Ser381 phosphorylation by cytoskeletal damage. (A) Effect of H-89 on YAP phosphorylation by cytoskeletal damage. NIH3T3 cells were pre-treated with 20 μM H-89 for 1 h followed by addition of latrunculin B or seeding onto poly-HEMA-coated dishes. (B) Effect of dnPKA on YAP phosphorylation by cytoskeletal damage. NIH3T3 cells were infected with Flag–dnPKA retroviruses. Selected cells were treated with latrunculin B or seeded onto poly-HEMA-coated dishes. (C, D) Effect of PKA agonist on YAP phosphorylation. NIH3T3 cells were stimulated with 20 μM forskolin and 500 μM IBMX for the indicated times. YAP phosphorylation (C) and localization (C, D) were determined. WCL, whole cell lysate; Nuc, nuclear lysate. Scale bar, 10 μm. (E) LATS activation by PKA agonist or cell detachment. LATS activation loop (AL) phosphorylation was determined in cells treated with the indicated stimuli. (F) Requirement of LATS1/2 for YAP phosphorylation by PKA agonist. Lats1−/−;Lats2fl/fl MEFs were transduced with either empty or Cre retroviruses. Selected cells were treated with forskolin/IBMX. *, nonspecific signal.