YM1/2 and Mac3 staining coincide in the allergic lung. A. Serial sections of lungs from a representative OVA/alum-primed mouse that received IL-4Rα+/+ BMM were stained with H&E, anti-F4/80, anti-Mac3, or anti-YM1/2 as shown. Digital images were taken from the same region of the lung at 40X and 100X. Arrow heads highlight the staining of representative macrophages in the airways, while the asterisk highlights staining of macrophages surrounding blood vessels. B. Co-expression by immunofluorescence microscopy. A representative lung section was stained with rat anti-Mac3 and rabbit anti-YM1/2 (or control serum), followed by goat anti-Rat-PE and goat anti-rabbit-FITC. Nuclei were stained with DAPI. Digital images were captured for each color and processed as described in Materials and Methods. The 3 color images were merged into a single image. Appearance of yellow indicates expression of both Mac3 and YM1/2. C. Five fields were analyzed for the total number of nuclei, total number of cells staining red only, number of cells staining green only, and number of cells with both colors (yellow). The percentage of cells in each category was calculated by dividing each number by the total number of nuclei per field. These results are representative of 2 independent experiments.