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. 2016 Oct 15;143(20):3686–3699. doi: 10.1242/dev.134023

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© 2016. Published by The Company of Biologists Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Fig. 3. — Glucocorticoid signalling is sufficient, but not essential, to specify alveolar fate. (A) Experimental design: Tomato⁺ E12.5 or 16.5 tip or stalk was grafted into E12.5 host lung and grown with 50 nM Dx throughout culture. (B) Examples of alveolar-fated tip grafts stained for: green, LPCAT1 (alveolar fate); red, Tomato (graft); white, PDPN (basal and AT1 cells). Arrowheads, PDPN⁺ AT1 cells. (C) Split bar graph showing results from B. Each type of graft was analysed in at least three independent experiments. (D) E12.5 wild-type lungs were grown with or without Dx for up to 6 days; two independent experimental replicates. Note precocious expression of alveolar markers in the presence of Dx. Lungs cultured without Dx do express LPCAT1 from experimental day 5. Green, LPCAT1 (late tip progenitors and type 2 cells); red, SOX9 (tip progenitors). (E,F) Sections of GR^−/− and GR^+/+ sibling lungs at E17.5 and E18.5 stained for: green, HOPX (AT1 cells); red, SOX9 (tip progenitors); white, E-CAD (epithelium) (E), and: green, LPCAT1 (late tip progenitors and AT2 cells); red, LAMP3 (AT2 cells); magenta, PDPN (late tip progenitors and AT1 cells) (F). A total of five GR^−/− and 5 GR^+/+ sibling lungs from three independent litters were observed at both E17.5 and E18.5. Blue, DAPI. Dashed line, edge of lung. Scale bars: 100 μm in B; 50 μm in D-F.