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. 2018 Jun 21;9:2424. doi: 10.1038/s41467-018-04663-1

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© The Author(s) 2018

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — Zeb2 blocks nuclear translocation of Yap1. a Immunostaining showing Yap1 is not concentrated in the nucleus in normal lung, AAH, and AD. b A cluster forming in the interior of an AD at P170. Arrows show the cluster border. c Yap1 is translocated to the nucleus in clusters. d The downstream target of Yap1, IL-6, is induced with nuclear translocation of Yap1 in clusters. e Cells in culture were allowed to reach confluence, and were then treated with 75 pM Tgf-β1 for 12 h. Note the block in Yap1 nuclear translocation at confluence. Similar cytoplasmic retention of Yap1 was evident at confluence in the absence of Tgf-β1. Zeb2 shRNA knockdown maintains nuclear Yap1 at confluence. f Real-time PCR showing Zeb1 mRNA is repressed and Zeb2 mRNA is induced when cells are allowed to become confluent. g Control cells remain growth-arrested at confluence, but knockdown of Zeb2 allows continued proliferation and cell outgrowth into foci. h Knockdown of Zeb1 leads to loss of proliferation and senescent morphology²⁹. i–i″ Yap1 is nuclear in subconfluent control cells. j–j″ Yap1 becomes sequestered in the cytoplasm of subconfluent cells when Zeb1 is knocked down. Bars are 50 μm in panels a–e, and 25 μm in panels f–j