Figure 2.

Sca-1 and CD34 expression following depletion of Clara cell secretory protein (CCSP)-expressing cells. (A): Flow cytometry analysis of Sca-1 and CD34 expression on the surface of CD45^neg CD31^neg live cells isolated from untreated mice (top row) or CCSP-HSVtk transgenic mice recovered for 6 days post-GCV treatment (bottom row). Left column represents unstained samples, and right column represents Sca-1 CD34 dual-stained sample. (B): Overlay of the autofluorescence (AF) profile (unstained sample) of CD45^neg CD31^neg live cells isolated from untreated mice (purple contour) or CCSP-HSVtk transgenic mice recovered for 6 days post-GCV treatment (black contour), demonstrating the disappearance of the AF^high population following ablation of CCSP-expressing airway epithelial cells. (C): Quantification of data in (A). The percentages of cells in each of the three gates (Sca-1^pos CD34^pos, Sca-1^neg CD34^pos, and Sca-1^pos CD34^neg) were compared between untreated (dark gray bars) and GCV-treated (light gray bars) samples. (D): Intracellular staining for CCSP of cells isolated from a wild-type animal showing the presence of CCSP-immunoreactive cells in the AF^high population. The panels represent unstained sample (top left), no primary control (top right), CCSP-stained sample (bottom left), and overlay of the AF profiles of the CCSP-positive (red contour) and no primary control (black contour plot) samples (bottom right). Abbreviations: GCV, ganciclovir; PE, phycoerythrin; Pe, phycoerythrin.