Multiorgan disease in mice with cGVHD; immune infiltration and collagen deposition. (A) Tissues (lung, liver, tongue, salivary glands [SG], thymus, spleen, ileum, and colon) were harvested at day 60 after transplantation and stained with H&E to determine pathology. Bright-field images were captured at 100× magnification with an Olympus BX51 microscope. (B) Inflammation, immune infiltration, and parenchymal changes were scored with a cumulative scoring system used previously.26 (C) Collagen deposition was determined with a Masson trichrome staining kit; blue indicates collagen deposition. (D) Collagen deposition was quantified as a ratio of blue area to total area of tissue. Quantification was performed with the analysis tool in Photoshop CS3. Data from 2 individual experiments were pooled to obtain pathology scores; n = 12. *P < .05; **P < .01; ***P < .005.