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. 2017 Jan 12;8(1):e2546. doi: 10.1038/cddis.2016.478

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Copyright © 2017 The Author(s)

Cell Death and Disease is an open-access journal published by Nature Publishing Group. This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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CRB3 is an upstream regulator of the Hippo pathway and affects FRMD6 transcription. (a) Protein levels of Hippo pathway components were examined in cells grown under sparse and confluent conditions. The numbers below the images indicate the relative expression levels of p-Lats1/Lats1 and p-YAP/YAP. (b) YAP localization was assessed in cells grown under sparse and confluent conditions. Scale bar, 25 μm. (c) Protein expression of the core regulators of the Hippo pathway was examined by immunoblotting. The numbers below the images represent relative expression levels of p-Lats1/Lats1 and p-YAP/YAP. (d-f) The expression levels of the FRMD6 and Kibra proteins and mRNAs were analysed by immunoblotting (d) and real-time PCR (e and f). ***P<0.001. Data are mean±S.D. (g) Cellular fractionation experiments were performed to examine the levels of the CRB3 protein in the nucleus and cytoplasm. GAPDH and Lamin A were used as loading controls. The numbers below the images indicate the relative expression level of YAP. (h) YAP localization was assessed in MCF10A cells grown under sparse and confluent conditions. Scale bar, 25 μm. (i, j) The expression levels of the CTGF and CYR61 mRNAs in MCF10A and T47D cells were examined using real-time PCR. shCRB3 cells were transfected with siRNAs to silence the expression of YAP. Cells were collected 48 h post-transfection. **P<0.01, ***P<0.001. Data are mean±S.D.