Fig. 5.

Three-tier orthogonal technology approach to the analysis of Mage-A4 expression in ER⁻ PgR⁻ Her2⁺ breast cancer tissue. A, Silver stained IEF 2D-PAGE analysis of DCTB patient 41 tumor sample (ER⁻ PgR⁻ Her2⁺), with MS identification of the Mage-A4 protein. Two spots were identified in this case. B, 2D Western blot analyses of Mage-A4 expression confirmed the presence of Mage-A4 (white arrows correspond to the MS-identified spots; antibody mAb 57B) and, as it was the case for TNBCs, revealed the existence of multiple forms of the protein (black arrowheads). C, Expression of the protein by tumor cells was confirmed by IHC (antibody mAb 57B; yellow arrow). Scale bar, 100 μm.