Fig. 1

Protocol of the flow cytometric analysis. Single lung cells were obtained from the mice and analyzed by flow cytometry. Most of the single lung cells, distributed according to their values of forward and side scatter, were gated at the initial window (the upper left panel). Next, doublets and dead cells were excluded by FSC-A/FSC-W and 7-AAD staining, respectively (the upper middle and upper right panels). The expression of the pan-leukocyte marker CD45 designated a hematopoietic fragment (fragment I). CD45-negative cells were further subjected to flow cytometric analysis for the expression of the endothelial cell marker CD105 and epithelial marker EpCAM. The cells were divided into the following fragments: the EpCAM⁺ CD105⁻ fragment (fragment II: epithelial cells), EpCAM⁻ CD105⁻ fragment (fragment III: mesenchymal cells) and EpCAM⁻ CD105⁺ fragment (fragment IV: endothelial cells). We classified single lung cells into the fragments I to IV according to this protocol and analyzed the cells for the presence or absence of GFP fluorescence in each fragment in subsequent experiments