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. 2017 Jul 4;8:15857. doi: 10.1038/ncomms15857

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Copyright © 2017, The Author(s)

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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Lentiviral-mediated transduction of HA-tagged E2F4 (diagrams: E2F4^WT, E2F4^ΔDBD, E2F4^ΔNES) or mCherry control constructs in E2f4^f/f;R26^CreERT2/+ airway progenitors treated with Tm (Tamoxifen). Left panels: qPCR of ALI day6: increased Foxj1 and Deup1 mRNAs by E2F4^WT or E2F4^ΔNES or but not E2F4^ΔDBD, compared to mCherry. Bars are mean (+s.e.m.). (a–c) Right panels: Immunofluorescence/confocal imaging: HA, acetylated α-tubulin, DAPI (diagram, apical and basal aspects). HA signals in both nucleus and cytoplasm in E2F4^WT and E2F4^ΔDBD but only in nuclei of E2F4^ΔNES. Rescue of multiciliogenesis (arrow) in E2f4-deficient cells by transduction of E2F4^WT but not E2F4^ΔDBD or E2F4^ΔNES (arrowheads: monocilia). Bar in a=5 μm.