Figure 6.

Roles of Src and FAK in the regulation of nuclear YAP by EGF or LPA. (A) Src depletion and FAK inhibition. MCF-10A cells transfected with control or Src siRNA were serum starved for 24 h and treated with EGF or LPA for 30 min. For FAK inhibitor treatment experiment, serum-starved, subconfluent MCF-10A cells were pretreated for 30 min with DMSO or 5 µM PF-573228, followed by a 30-min EGF or LPA treatment. YAP subcellular localization was determined by immunofluorescence staining. One of three independent results is presented. (B) CSK expression to inhibit Src activity. Paired with uninduced cells, doxycycline (Dox)-induced MCF-10A cells expressing myr-CSK-GFP fusion protein were serum starved for 24 h. Cells were treated with EGF or LPA for 30 min before fixation. Subcellular localization of YAP was determined by immunofluorescence staining. One representative out of three independent experiments is shown. Tet, tetracycline (or doxycycline) inducible. (C) Src inhibitors. Serum-starved, subconfluent MCF-10A cells were pretreated with PP3, PP2, imatinib, or dasatinib for 30 min. Cells were then treated with 20 ng/ml EGF for 30 min before fixation. Endogenous YAP was immunofluorescence stained. The result represents three independent experiments. Bars, 25 µm.