Extended Data Fig. 8: LGR5-mRFP reporter validation and co-culture.
From: Human distal lung maps and lineage hierarchies reveal a bipotent progenitor
a. Schematic depicting pLKO-LGR5-mRFP reporter construct. b. Representative gating for FACS sorting of LGR5-mRFP reporter fibroblasts used for organoid co-cultures. c. RNA-FISH for LGR5 (left), and PDGFRα (right) on LGR5-mRFP fibroblasts. d. Relative mRNA expression of LGR5, WNT5A, WNT2, CA3, and FGF7 in cultured LGR5-mRFP fibroblasts vs alveolar fibroblasts. Data are presented as mean ± SEM. (** p = 0.0055 for LGR5), (* p = 0.0187 for WNT5A), (NS p = 0.0972 for CA3), (NS p = 0.1216 for WNT2), and (NS p = 0.1525 for FGF7) (unpaired two-tailed t-test with Welch’s correction) (n = 3). e. Immunostaining for SFTPB (green), TP63 (red) and KRT5 (grey) in basal cell organoid co-cultured with LGR5-mRFP fibroblasts or alveolar fibroblasts. blue (DAPI) indicates nuclei. Scale bar: 50 µm. f. Relative mRNA expression of SCGB3A2 and SOX2 in AT2s co-cultured with LGR5-mRFP fibroblasts. Data are presented as mean ± SEM. NS p = 0.2383, and *p = 0.0383 and (unpaired two-tailed t-test with Welch’s correction) (n = 3).
