Figure 2.

Analysis of bleomycin-induced inflammation in TSP-1^−/− mice. (A) TSP-1^−/− and WT mice suffer inflammation after bleomycin injection. Representative hematoxylin-and-eosin stains of these lungs are shown. Data are representative of six mice per group. (B) TSP-1^−/− mice tended to exhibit more inflammation after bleomycin challenge. A certified veterinary pathologist blindly scored each slide for inflammation. Shown is the average pathology score ± SEM for six mice per group. No inflammation was evident in vehicle-treated control mice (data not shown). (C) Analysis of cellular recruitment. Slides were subjected to immunohistochemical analysis. From each slide, 10 images were obtained using an Olympus microscope with digital camera (Center Valley, PA). Average numbers of pixels corresponding to specific stains were quantitated using Adobe Photoshop software. Average data ± SEMs are shown for a minimum of five animals per treatment group. All P values not shown are > 0.20. (D) Fresh lungs were flushed with 1.5 ml of sterile PBS. Cell pellets were obtained and resuspended in 200 μl PBS, and cells were counted using a hemacytometer. Cells in PBS were cytospun and stained. One hundred cells were counted to obtain cell differential percentages, and total cell-line numbers were obtained by multiplying the differential by the total cell count. Macs, macrophages; Lymphs, lymphocytes; Neuts, neutrophils.