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. 2020 Mar 10;39(18):3754–3773. doi: 10.1038/s41388-020-1251-2

Fig. 1. CSC-like property and tumor initiation capacity of lung progenitor AT2 cells require interaction with Lgr5 cells in Gprc5a-knockout mice.

Fig. 1

IF analysis of the expression and localization (a) and differences (b) of AT2 cells (marked by surfactant protein A, SPA) in the S/TB region in 6-month aged Gprc5a—knockout (KO) or wild-type (WT) mice. Bar = 100 µm; c–g Isolated AT2 cells (Sca-1+CD31−CD45−) (c), Lgr5 cells (d) from 6-month aged mice, coculture of AT2 and Lgr5 cells (e) and colony formation analysis (f). Bar = 500 µm; Comparison of colony formation capacity of AT2 cells isolated from WT or KO mice cocultured with Lgr5 cells isolated from WT or KO mice (g); h Flowchart of the isolation and culture of Lgr5 cells from 6-month aged WT or KO mice and collection and centrifugation of supernatant; i Supernatant from Lgr5 cells applied in a 3D culture system of AT2 cells; j Observation of spheroids of AT2 cultured with supernatant from Lgr5 cells (left) (Bar = 1000 µm) and a higher magnification image (right); k Comparison of the spheroid formation capacities of AT2 cells isolated from 6-month aged KO mice and cultured with the supernatant from Lgr5 isolated from WT or KO mice at same age; Illustration (l) and comparison of the tumor initiation capacities (m) of isolated AT2 and Lgr5. KO:AT2 cells cultured with the supernatant of KO:Lgr5 or WT:Lgr5 cells or cocultured with KO:Lgr5 or WT:Lgr5 cells for 72 h and subcutaneously injected into mice to evaluate the tumor initiation capacity. Data were collected from three independent experiments with triplicate samples. ***P < 0.001.