Xenbase Image ID: 153510

Figure 3. Hh/Gli Signaling Is Required for Mouse Respiratory Specification (A–F) Immunofluorescence of control and Gli2;Gli3 mutant mouse embryos at E9.5 (23–26 s) in whole mount (A–D) and sections (E and F) reveal that Gli2 and Gli3 are required for Nkx2-1+ respiratory progenitors (thyroid, th; lung, lu; liver, li; pancreas, pa; hindgut, hg; and neural tube, nt). (G) In situ hybridization and immunostaining of E9.5 (23–27 s) sections show that Wnt2, Wnt2b, and Bmp4 expression, as well as pSmad1 require Gli2 and Gli3. The yellow dotted line outlines the foregut epithelium, and the red dotted line indicates the splanchnic lpm. (H and I) The average number of foregut cells (H), as well as the average % of mitotic (phospho-Histone H3+) and apoptotic (cleaved caspase-3+) foregut cells (I) was quantified from immunostained sections (n = 3 sections/embryo) of control and Gli2;Gli3 mutant embryos at E8.5 and E9.5 (n = 3 embryos each). The total cell number was based on DAPI staining (±SD, ∗p < 0.05 in pairwise Student’s t test of mutant and age matched controls). (J–M) Treatment of E8.5 mouse foregut explants with cyclopamine (cyclo) for 2 days results in loss of Nkx2-1+ respiratory progenitors phenocopying the Gli2−/−;Gli3−/− mutants. Nkx2-1+/Foxa2+ respiratory fate in Hh-signaling deficient explants was rescued by addition of CHIR (to stabilize β-catenin) and BMP4 (thyroid, th, and lung, lu). See also Figure S3. Image published in: Rankin SA et al. (2016) Copyright © 2016. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Larger Image Printer Friendly View

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