Fig. S5

Pathology in adult livers. Low magnification views of wildtype (a) and mutant (c, e) livers stained with hematoxylin and eosin. Scale bars are 200 μm. Lesions (arrows) are present in 1 of 4 wildtype and 4 of 4 mutant livers analyzed (g). Each liver is outlined by dashed lines. Magnified views of the same livers (b, wildtype; d, f, mutants). Scale bars are 40 μm. a, Small lesions (70–160 μm in diameter) are scattered throughout the liver parenchyma in one of four wildtype livers. b, An example of one of the lesions found in the wildtype liver. They appear to be well-circumscribed lesions composed of approximately 1–3 layers of flattened epithelioid cells (inset) surrounding eosinophilic material containing occasional golden-brown pigment granules and fragments of nuclear debris. Minimal inflammatory cells are present around the lesions and the hepatic parenchyma appears normal. c, An example of the mutant livers which show abundant and larger lesions (arrows and arrowhead). d, Magnified view of the nodule marked by the arrowhead in c. These lesions have a more exuberant epithelioid proliferation at the periphery, often more than 10 cell layers thick, and contain degenerating cellular debris, golden-brown pigment (arrows), and scattered calcifications. Focal inflammation surrounds a few lesions but they are, for the most part, associated with minimal adjacent inflammatory reaction. The hepatic parenchyma of the mutants is unremarkable in some areas but in other areas shows varying degrees of disorganization. e, Mutant #3 shows abundant and occasionally coalescing lesions (arrowhead). f, A magnified view of the coalesced lesion shows calcified material in the center (*). The hepatic parenchyma in mutant #3 shows hepatocyte dropout and increased fibrous tissue surrounding several of the lesions. g, Plot of the diameter of each lesion found in each liver. The bar indicates the average lesion diameter. Three of the four wildtype livers did not present with lesions. The plot shows that the severity of the phenotype varies in mutants but that on average, the lesions are almost twice to three times the size of those in wildtype #3. Mutant #3, the most severely affected, presented with 74 lesions of varying diameters.