Immunohistochemical analysis showing loss of Melan-A, but not CT antigen expression over time. Staining for Melan-A (antibody M27C10), NY-ESO-1/LAGE-1 (antibody D8.38, a gift from G. Spagnoli, Basel, Switzerland), and MAGE-A proteins (detected either by clone 57B, top 2 panels, or clone 6C1, bottom 2 panels) was performed on paraffin sections of metastases removed at the times indicated. Subcutaneous metastases (Apr. 03, Mar. 04), LN (Oct. 03) and the last brain metastasis (June 07) are shown. Insets show CT antigen negative areas in the last metastasis.