Fig. 3: Functional analysis of human airway basal progenitor cells in vivo.
From: Cloned airway basal progenitor cells to repair fibrotic lung through re-epithelialization
a Spatial visualization of the mouse (red) and human (green) epithelial cell marker CDH1 (E-cadherin) and CLDN4 gene expression in different lung zones. Yellow circle: damaged honeycombing zone; red circle: normal alveolar tissue zone; green circle: saccular zone; blue circle: pod zone. Scale bar, 1 mm. b Spatial visualization of mouse fibrotic marker Collagen1 (Col1a1) gene expression in different lung zones. High magnification indicated the Collagen1 signal in pod and saccule zones. Scale bar, 1 mm. c Expression level of multiple mouse marker genes in different lung zones. Normal, normal alveolar area; Damage, damaged honeycombing zone; Sac, saccular zone (n = 5 technical replicates). d Immunofluorescence images of mouse capillary vessels in the saccular zone with anti-GFP (green), anti-CD31 (red), and anti-CD34 (red) antibodies co-staining and DNA counterstain (DAPI, blue). Scale bar, 50 μm. e Barrier function assays of airway basal progenitor cells in vitro. Left, schematic diagram of the barrier function transwell assays; middle, TEER assay of monolayers formed by airway basal progenitor cells or A549 cells before and after Triton X-100 treatment (n = 6 biological replicates); right, assay of fibroblast migration through monolayers formed by airway basal progenitor cells or A549 cells (n = 6 biological replicates). f Quantification of the inflammatory area of mouse lung with or without airway basal progenitor cell transplantation (n = 8, 12, 15 technical replicates from 3 biological replicates). g Ashcroft score of lung fibrosis and quantification of the relative fibrotic area by Masson’s trichrome staining of mouse lungs with or without airway basal progenitor cell transplantation (n = 8, 12, 15 technical replicates from 3 biological replicates). h O2 partial pressure to CO2 partial pressure ratio of mouse arterial blood with or without human airway basal progenitor cell transplantation (n = 9 biological replicates). i Survival curve of mice challenged by bleomycin with or without airway basal progenitor cell treatment (n = 3, 4, 5 biological replicates). Statistical analysis was performed using the Log-rank (Mantel-Cox) test. c, e–h All data are presented as mean ± SEM. Statistical analysis was performed using one-way ANOVA with Tukey’s post hoc test and two-way ANOVA with Sidak’s multiple comparisons test (for the middle panel of E) (**P < 0.01, ***P < 0.001, and ****P < 0.0001; ns, no significant difference).
