Fig. 5.

Increased cell proliferation and reduced AECII senescence in p16^−/− lungs. a Twenty-four hours post IP EdU injection (50 mg/kg), p16^+/+ and p16^−/− lungs exposed to RA and CS were fixed and stained with Click-It EdU (green), SPC (yellow), Podoplatin (PDPN, red), and DAPI (blue, scale bar = 30 µm). b Quantification of lung proliferating (EdU+) cells in the alveolar airspace. At least 1500 nuclei per airspace were counted in each lung *p = 0.0197, **p < 0001. N = 4–12 lungs per group. c Percentage of SPC+ cells in the alveolar region of the lung *p = 0.015, **p = 0.0004. d Images of Isolated AECIIs from p16^+/+ and p16^−/− lungs treated with 5% CSE in 10% serum for 24 h and probed for β-galactosidase activity using C12FDG substrate (scale bar = 400 µm). e AECIIs immunostained with SPC (red) and DAPI (blue). f Flow cytometry analysis of AECII measuring purity of isolation. X axis is measuring EPCAM for epithelial cell specificity and y axis is measuring SPC for AECII specificity. g AECIIs treated with 5% CSE and C12FDG, and quantified with flow cytometry. Data represent the average ± SEM of four to five independent experiments