Fig. 5: The number of mito-ncR-805 spots is decreased in AETII cells of mice acutely and chronically exposed to CS.

a–d Mice were exposed to smoke from one cigarette (a) or to smoke from 2 cigarettes per day, 5 days a week, for 6 months (b–d). Lungs were inflated, embedded into paraffin blocks, 4 micron sections were cut, and analyzed. a, d Expression of mito-ncR-805 (blue) and Surfactant B mRNA (red) using ISH. Images were acquired using ×100 objective. Representative images are shown, scale bar is 20 μm. Arrows indicate enlarged AETII cells shown in (1) and (2) panels, respectively. Scale bar for enlarged images is 5 μm. Hoechst counterstain overlay is removed from enlarged images to maximize contract of mito-ncR-805 and Surfactant B signals. b Examples of lungs of sham and mice exposed to CS for 6 months stained with Gill staining to visualize alveolar compartment. Scale bar is 100 μm. c Morphometric analysis of average alveolar chord length of sham and 6-month CS-exposed mice, where sham n = 6 animals and 6-month CS n = 9 animals. e Quantification of the number of mito-ncR-805 spots in AETII and non-AETII cells. Each point represents counting from 6 to 8 randomly chosen fields acquired at ×100 magnification from each individual animal. For controls, n = 76 AETII and 480 non-AETII cells. For 12 h of CS exposure, n = 79 AETII and 403 non-AETII cells. For 24 h, n = 87 AETII and 472 non-AETII cells. For 48 h, n = 53 AETII and 376 non-AETII cells. For the sham group, n = 61 AETII and 322 non-AETII cells. For chronic smokers, n = 64 AETII and 414 non-AETII cells. P values are pControl^{AETII/non-AETII} = 5.55558E–08, p^{Control/12h-AETII} = 1.04561E–18, p^{Control/12h-non-AETII} = 0.004329157, p^{Control/24h-AETII} = 1.43455E−13, p^{Control/24h-non-AETII} = 0.002372331, p^{Control/48h-AETII} = 3.89362E–11, p^{Control/48h-non-AETII} = 0.054832469, p^{Sham/6 mo-AETII} = 9.28908E−21, p^{Sham/6 mo-non-AETII} = 0.000104857, p6-mo-sm^{AETI/non-AETII} = 0.057089623.