Fig. 5

AT2 cell isolation in an LPS-induced lung injury model. a Hematoxylin and eosin staining of the lung at 24 h after intratracheal instillation of LPS (1 μg/g body weight). Alveolar spaces were infiltrated with inflammatory cells. Scale bars, 50 μm. b Representative FACS plot that identifies CD45⁻CD31⁻ cells (left), which were analyzed for EpCAM and MHCII expression to sort P1 cells (right) in control samples. c Representative FACS plots of the LPS-induced lung injury model. Compared to the control, the proportion of CD45⁻CD31⁻ cells was lower due to increased numbers of inflammatory cells (left). P1 cells were successfully identified and sorted (right). In both the control and the LPS model, whole-lung samples and sorted P1 cells were analyzed for (d) Sftpc, (e) Cxcl1, (f) Tnf, (g) Mmp2, (h) Mmp9, and (i) Mmp12 mRNA expression (n = 3/group). In P1 cells, Cxcl1, Tnf, and Mmp9 were upregulated following LPS stimulation. Mmp2 and Mmp12 were undetected in P1 cells. EpCAM, epithelial cell adhesion molecule; MHCII, major histocompatibility complex class II; PE-Cy7, phycoerithrin-cyanin7; APC, allophycocyanin; LPS, lipopolysaccharide; SSC, side scatter