Fig. 2: Characteristics of MEF-derived iPULs.
From: Direct reprogramming of mouse fibroblasts into self-renewable alveolar epithelial-like cells
a Schematic showing the passaging and cryopreservation of iPULs. b mRNA expression levels of the indicated genes in MEFs, unpurified iPULs (8 days after transduction), purified iPUL (P6–7) organoids, AT2 organoids (P3–6), primary AT2 cells isolated from Sftpc-GFP mice (8–18 weeks old), whole adult lungs (12–18 weeks old), and fetal lungs (E18.5). Data are presented as mean values ± SEM; n = 3 biologically and technically independent samples. c Representative hematoxylin and eosin (H & E)-stained images, GFP luminescence (green), and immunofluorescence staining for pro-Sftpc (AT2 cell marker, red) and Ager (AT1 cell marker, gray) in iPUL organoids. Scale bar: 50 µm (H & E) and 100 µm (others). d Transmission electron microscopy images of iPUL organoids (P6) containing lamellar bodies and microvilli (white arrows). Scale bar: left = 5 µm and right = 500 nm. e Live cell imaging of iPUL organoids at the indicated time points after the addition of LysoTracker Red DND-99 (100 nM) to the medium. Scale bar: 100 µm. f Principal component analysis (i), row-normalized heatmap data, and hierarchical clustering (ii) based on the RNA-seq analysis of MEFs, unpurified iPULs (8 days after transduction), purified iPULs (P6–7), AT2 organoids (P3–6), primary AT2 cells isolated from Sftpc-GFP mice (8–18 weeks old), adult lungs (12–18 weeks old), and fetal lungs (E18.5).
