FIGURE 4.

GPCR agonists and serum induce cytoplasmic localization and phosphorylation of YAP in IEC-6 and Caco-2 intestinal epithelial cells. A–D, upper, confluent cultures of IEC-6 (A), Caco-2 (B), HEK293 (C), and PANC-1 (D) cells were stimulated with 10 nm ANG II (A), 10% FBS (B and C), or 10 nm neurotensin (NT) (D), as indicated. The cultures were then washed, fixed with 4% paraformaldehyde, and stained with an antibody that detects total YAP and with Hoechst 33342 to visualize the cell nuclei. A–D, lower, confluent cultures of IEC-6 (A), Caco-2 (B), HEK293 (C), and PANC-1 (D) cells were stimulated with 10 nm ANG II (A), 10% FBS or 10 μm lysophosphatidic acid (LPA) (B and C), or 10 nm neurotensin (NT) (D) for the indicated times. Cultures were then lysed with 2× SDS-PAGE sample buffer and analyzed by immunoblotting antibodies that detect YAP phosphorylated at Ser¹²⁷ and total YAP. Similar results were obtained in at least two independent experiments.