Volume 50, Issue 5, Pages (November 2006)

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Volume 50, Issue 5, Pages 1102-1110 (November 2006) Mitogen-Activated Protein Kinase Signaling is Activated in Prostate Tumors but not Mediated by B-RAF Mutations Maximilian Burger, Stefan Denzinger, Christine Hammerschmied, Andrea Tannapfel, Armin Maderstorfer, Wolf Ferdinand Wieland, Arndt Hartmann, Robert Stoehr European Urology Volume 50, Issue 5, Pages 1102-1110 (November 2006) DOI: 10.1016/j.eururo.2005.11.031 Copyright © 2006 European Association of Urology Terms and Conditions

Fig. 1 (A) Specificity of allele-specific PCR (PASA) for detection of the V600E mutation of the B-RAF gene. Lanes 1 and 2, positive controls (colon cancer cell line HT29 and melanoma cell line SK-MEL3); lanes 3 and 4, negative controls (bladder cancer cell line RT4, colon cancer cell line SW480); lane 5, H2O. (B) Representative PASA screening of 6 prostate tumors (lanes 1–6). Lane 7 shows V600E polymerase chain reaction (PCR) product from the HT29 cell line (positive control); in lane 8 RT4 DNA was used as negative control (lane 9, H2O). None of the analyzed tumors showed the mutation-specific PCR product (lower lane). The upper lane shows PASA for the wild-type V600 allele. The presence of a PCR product indicates a sufficient amount and quality of the DNA for analysis. (C) Microsatellite instability (MSI) analysis of a prostate tumor using the marker BAT40: NO, normal prostatic tissue; TU, prostate tumor. Extensive band shift indicated by arrows can be seen in the tumor (≥MSI positive). (D) Immunohistochemical staining of 2 prostate tumors using an antibody specifically recognizes the dually phosphorylated active form of mitogen-activated protein kinase (MAPK). Tumor cells show strong staining indicating active MAPK signaling (magnification: ×200). European Urology 2006 50, 1102-1110DOI: (10.1016/j.eururo.2005.11.031) Copyright © 2006 European Association of Urology Terms and Conditions