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. 2013 Mar;48(3):288–298. doi: 10.1165/rcmb.2011-0403OC

Figure 4.

Figure 4.

Characterization of GFP-labeled cells in postnatal and adult lungs. (A) Bar graph shows the percentage of SPC+ cells with SPC immunoreactivity in alveolar regions of variously aged SPC H2B-GFP mice. Twenty alveolar regions from four lung sections per lung and three mice for each age were scored by IF with antibodies for SPC and GFP. P, postnatal. (B) IF in sectioned lungs from 8-week-old SPC H2B-GFP mice shows GFP expression in terminal bronchiolar cells, involving CCSP (purple), SPC (red), and GFP (green). Boxed areas are magnified in C–E. Scale bar, 100 μm. (C) Terminal bronchioles retain weak GFP expression. (D) GFP-labeled cells coexpress SPC and CCSP in bronchioalveolar duct junctions. (E and F) IF with antibodies for GFP, SPC, and CCSP (E) and for GFP and calcitonin gene-related peptide (red) (F) in serial sections shows that GFP-labeled cells reside around pulmonary neuroendocrine cells. (G) Bar graph shows the proportion of GFP-labeled cells with the indicated patterns of CCSP and/or SPC immunoreactivity at various ages in SPC H2B-GFP mice. Twenty alveolar regions, 20 bronchioles, and 20 terminal bronchioles with 4–8 lung sections per lung from three mice for each age group were scored by IF with antibodies for SPC, CCSP, and GFP. (H–I) Quantitative PCR for assessing the expression of lung markers SPC (H) and CCSP (I) in GFP− versus GFP+ cells from variously aged SPC H2B-GFP mice. The y axis shows relative quantifications normalized to GAPDH. The data are representative of two independent experiments.