Perlecan quantification in metastatic nodules by immunohistochemistry. Staining melanocytic samples with Giemsa results in a uniform dark green/blue coloration of melanin that is easily distinguishable from the DAB precipitate of the HSP reaction. DAB-specific threshold selection from selected tumor areas was performed by ImageJ software (red/orange selection in Figures A, B, C and D), and total selected area was quantified and statistically analyzed (E). a: normal mice; b: untreated mice with B16F10 lung metastasis; c: vehicle-treated mice with B16F10 lung metastasis and d: M8-treated mice with B16F10 lung metastasis. M8 treatment resulted in a statistically significant decrease in heparan sulfate proteoglycan expression. A = Normal lung; B = B16F10 positive control; C = Vehicle treatment; D = M8 treatment. Y axis of graphs = transformed mean after normal distribution using the equation to transform data, .