Fig. 5

Functional and quantitative assessment of mitochondria in mouse tracheal and lung epithelium. MTECs and lung cells were collected from WT, Aldh2*2 Tg, and Aldh2 ^−/− mice, n = 4-5 mice/genotype. Cells were immunostained for the mitochondrial membrane protein, Tim23. No difference in fluorescence intensity was observed between WT (a) and Aldh2*2 Tg (b), but MTECs from Aldh2 ^−/− mice showed significantly lower fluorescence intensity (c). Quantification of fluorescence intensity from multiple cells from each genotype is shown in (d). Cells were also stained for MitoTracker (MT) probes and examined by fluorescence microscopy and flow cytometry. In comparison to WT cells (e), lung cells from Aldh2*2 Tg mice (f) had a higher fluorescence intensity, while the Aldh2 ^−/− mice (g) showed lower fluorescence intensity. Quantification of fluorescence intensity from multiple cells from each genotype is shown in (h). Not Aldh2*2 Tg (i) but Aldh2 ^−/− (j) showed a higher level of mitochondrial ROS than WT mice did. MTECs from WT and Aldh2 ^−/− mice were examined using a flux analyzer to measure and compare OCR (k) and ECAR (l) as indicators of the cellular mitochondrial respiration and glycolysis. No significant difference was observed between WT and Aldh2 ^−/− cells. a-c, e-g scale bars: 10 μm, nuclei were stained with DAPI (blue)