Fig. 1
Characterisation of pulmonary pericytes during alveologenesis. a, b Three-dimensional reconstruction (a) and high magnification thin optical section (b) of confocal images with Airyscan detection showing AQP5-stained type 1 alveolar epithelial cells (green), PDGFRβ-stained pulmonary pericytes (PCs) (red) and PECAM1-stained endothelial cells (ECs) (blue) in lung at 4 weeks. Panels on the right show higher magnification of corresponding insets (a) or single channels (b). Scale bar, 30 µm (a, left), 15 µm (a, right), 10 µm (b, left) and 5 µm (b, right panels). c, d Three-dimensional reconstruction (c) and thin optical section (d) of confocal images showing SFTPC-stained, cuboidal AT2 cells (green), PDGFRβ-stained PCs (red) and RAGE-stained AT1 cells (blue) in lung at 4 weeks. Panels on the right show higher magnification of corresponding insets. Scale bar, 20 µm (left), 10 µm (right). e Quantitation of EC to PC ratio as measured in peripheral lung sections at different developmental stages. Data represents mean ± s.e.m. (n = 5 for P7, n = 6 for P21 and Adult mice). f Scheme showing the time points of tamoxifen administration (P1–3) and analysis for the Pdgfrb(BAC)-CreERT2 R26-mT/mG mice. g–k High magnification images of Pdgfrb(BAC)-CreERT2 R26-mT/mG lung sections at indicated stages showing pulmonary GFP+ PCs (green), PECAM1+ ECs, PDGFRβ+ PCs, PDGFRα+ fibroblasts or αSMA+ bronchial smooth muscle cells/myofibroblasts (red, as indicated). Arrows in g indicate PC cell bodies, in g–k GFP-positive PDGFRβ+ cells (g, h), GFP-negative PDGFRα+ cells (i) or αSMA+ cells (j, k) are marked. Scale bar, 20 µm (g, h), 15 µm (i, k) and 30 µm (j). l RT-qPCR analysis of Pdgfrb (pericytes), Pecam1 (endothelium), and Sftpc (epithelium) expression in freshly sorted GFP+, CD31+ or EpCAM+ cells from P7 Pdgfrb(BAC)-CreERT2 R26-mT/mG lung. Data represents mean ± s.e.m. (n = 4 mice)
