bioimagesUnknownXia GaoThe Journal of Cell Biologyhttps://www.ebi.ac.uk/biostudies/studies/S-JCBD-201506014JCB

Sections of spheres stained for Grhl2 (red), Krt8 (green), Pdpn (BC marker, magenta) and DAPI (blue).

IHC of section through human lung (6 mm diameter airway) for SMAGP (red), Foxj1 (green) and DAPI (blue).

Representative DIC and fluorescence microscopy images at culture day 14 of spheres formed in Matrigel from lineage-labeled Ngfr+ BCs isolated by FACS (red) from tracheas of Krt5-CreER; Grhl2 flx/-; Rosa-tdTm mice.

Immunohistochemistry for Smgap (red) and Trp63 (green) of wild type spheres.

Immunohistochemistry for ZNF750 (red) and Trp63 (green) of wild type spheres.

Representative DIC and fluorescence microscopy images at culture day 14 of spheres formed in Matrigel from lineage-labeled Ngfr+ BCs isolated by FACS (red) from tracheas of Krt5-CreER; Grhl2 flx/+; Rosa-tdTm mice.

At day 21, spheres were fixed, sectioned and stained for SMAGP (red), a-tubulin (green) and DAPI (Blue).

Representative DIC and fluorescence microscopy images at culture day 14 of spheres formed in Matrigel from lineage-labeled Ngfr+ BCs isolated by FACS (red) from tracheas of Krt5-CreER; Rosa-tdTm mice.

Immunohistochemistry for ZNF750 (red) and Trp63 (green) of GRHL2 mutant spheres.

At day 14, spheres were fixed, sectioned and stained for (A) Grhl2 (red), Trp63 (green) and DAPI (blue).

Wholemount staining of tracheal epithelium of Krt5-CreER; Rosa-tdTm and Krt5-CreER; Grhl2flx/-; Rosa-tdTm mouse for RFP (red), Scgb1a1 (green), a-tubulin (magenta) and DAPI (blue) 10 d after SO2 inhalation. Images are maximum intensity projections generated from z stack confocal images.

Sections stained for Scgb3a2 (red), Foxj1 (green) and DAPI (blue).

IHC of tracheal sections stained for a-tubulin, Scgb1a1, RFP and DAPI.

At day 14, spheres were fixed, sectioned and stained for (A) Grhl2 (red), Foxj1 (green) and DAPI (blue).

Sections of spheres stained for E-cadherin (E-cad, green), Cldn4 (red) and DAPI (Blue).

IHC of section through human lung (6 mm diameter airway) for SMAGP (red), Trp63 (green), and DAPI (blue).

IHC of section through human lung (6 mm diameter airway) forZNF750 (red), Trp63 (green) and DAPI (blue). Single images from confocal stacks.

At day 21, spheres were fixed, sectioned and stained for SMAGP (red), E-cadherin (green) and DAPI (Blue).

Higher magnification of the boxed region.

Immunohistochemistry for Smgap (red) and Trp63 (green) of GRHL2 mutant spheres.

At day 21, spheres were fixed, sectioned and stained for ZNF750 (red) and a-tubulin (green).

IHC of section through human lung (6 mm diameter airway) forZNF750 (red), Foxj1 (green) and DAPI (blue). Single images from confocal stacks.

At day 21, spheres were fixed, sectioned and stained for GRHL2 (red), Foxj1 (green) and DAPI (blue)

bioimagesImage 632922 (Figure 4 - B)Figure 8 - AFigure 8 - BImage 632917 (Figure 4 - A)Figure 7 - DImage 632937 (Figure 8 - B)Image 634627 (Figure 5 - F)Image 634623 (Figure 4 - D)Image 632945 (Figure 3 - A)Image 632934 (Figure 7 - D)Image 632986 (Figure 2 - C)Image 632946 (Figure 3 - A)Image 632932 (Figure 6 - D)Image 632912 (Figure 2 - B)Image 632943 (Figure 6 - D)Image 632938 (Figure 8 - B)Figure 2 - CImage 632931 (Figure 6 - D)Image 632913 (Figure 2 - B)Image 632936 (Figure 8 - A)Figure 2 - BImage 632944 (Figure 3 - A)Image 632942 (Figure 6 - D)Image 632985 (Figure 2 - C)Image 632916 (Figure 4 - A)Image 634633 (Figure 6 - D)Figure 3 - AImage 632935 (Figure 8 - A)Figure 4 - AFigure 4 - BImage 632923 (Figure 4 - B)Image 632920 (Figure 4 - A)Image 634630 (Figure 5 - G)Figure 4 - CFigure 4 - DImage 632927 (Figure 4 - C)Image 634626 (Figure 5 - F)Image 634628 (Figure 5 - G)Image 632933 (Figure 7 - D)Image 632925 (Figure 4 - B)Figure 5 - FImage 634629 (Figure 5 - G)Image 632919 (Figure 4 - A)Figure 5 - GImage 632921 (Figure 4 - A)Image 634631 (Figure 5 - G)Figure 5Image 634632 (Figure 6 - D)Figure 4Image 632928 (Figure 4 - D)Figure 7Figure 6Image 632926 (Figure 4 - C)Figure 8Image 632918 (Figure 4 - A)Figure 6 - DImage 632924 (Figure 4 - B)Figure 3Figure 2Xia GaoAman S. BaliScott H. RandellBrigid L.M. HoganfalseGRHL2 coordinates regeneration of a polarized mucociliary epithelium from basal stem cells<br /> Pseudostratified airway epithelium of the lung is composed of polarized ciliated and secretory cells maintained by basal stem/progenitor cells. An important question is how lineage choice and differentiation are coordinated with apical–basal polarity and epithelial morphogenesis. Our previous studies indicated a key integrative role for the transcription factor Grainyhead-like 2 (Grhl2). In this study, we present further evidence for this model using conditional gene deletion during the regeneration of airway epithelium and clonal organoid culture. We also use CRISPR/Cas9 genome editing in primary human basal cells differentiating into organoids and mucociliary epithelium in vitro. Loss of Grhl2 inhibits organoid morphogenesis and the differentiation of ciliated cells and2015-11-02T11:28:01Z2018-11-29T11:28:01Z2018-11-29T11:28:01ZS-JCBD-20150601410.1083/jcb.201506014