Fig. 2
Establishment and characterization of FTE organoids a Bright field images of organoid developing from a single FTE cell after the indicated days of culture. Scale bar, 20 μm. b Immunofluorescence staining for ciliated cell marker acetyl-α-tubulin (green) and secretory cell marker PAX8 (red) in FTE organoid after 7 days in culture. Scale bar, 2 μm. c Top: Schematic showing generation of FTE organoids from PTPT mice. Bottom: GFP in organoids with or without Dox treatment; Dox (500 ng/ml) was added after seeding, and GFP− cells were removed by FACS 4 days after Dox addition. The organoid in the right panel was photographed before Dox addition; the organoids in the left panel were photographed after Dox (see the “Methods” section for details). Scale bar, 10 μm. d Sections of PTPT organoids after two consecutive passages, with or without Dox treatment (500 ng/ml), stained with H&E or subjected to immunofluorescence or IHC staining for the indicated markers. Scale bar, 20 μm. e Organoids established from the indicated mice, incubated with EdU (2 μM) and DAPI (1 μg/ml) for 2 h on day 7 of culture and visualized by immunofluorescence; scale bar, 20 μm. f % EdU-positive cells in organoids established from the indicated mice. g Representative immunofluorescence images of PAX8 (red) and acetylated-α-tubulin (green) in the indicated organoids at day 7 of culture; scale bar, 20 μm. h % ciliated cells (aceylated-α-tubulin+) in organoids established from the indicated mice. i Representative images of the bottom surface of Transwell units seeded with the indicated organoids. j Quantification of invasive cells in i. Data represent mean ± SEM from three mice of each genetic background. *P < 0.05, **P < 0.01,***P < 0.001, Tukey’s multiple comparison test. Source data are provided as a Source Data file.
