Discrimination of Melanocytic Tumors by cDNA Array Hybridization of Tissues Prepared by Laser Pressure Catapulting Bernd Becker, Alexander Roesch, Christian.

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Discrimination of Melanocytic Tumors by cDNA Array Hybridization of Tissues Prepared by Laser Pressure Catapulting Bernd Becker, Alexander Roesch, Christian Hafner, Wilhelm Stolz, Michael Landthaler, Thomas Vogt Journal of Investigative Dermatology Volume 122, Issue 2, Pages 361-368 (February 2004) DOI: 10.1046/j.0022-202X.2004.22240.x Copyright © 2004 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 1 (a) A typical protocol during SMART cDNA amplification PCR with two different templates recorded with the Rotor-Gene real-time PCR machine. The marks A and B label the number of cycles used for equal amplification of the templates; 22 and 25 cycles, respectively. -RT represents the control reaction without reverse transcriptase during the first-strand synthesis step in the SMART protocol. The inset shows an agarose gel with the cDNA isolated from template A after 22 cycles and template B after 25 cycles; on the lane labeled -RT the negative control is shown. Equal volumes from the cDNA and the negative control were loaded on each lane. M: DNA size marker (sizes given in kb). (b) Array comparison dotplot of two arrays hybridized with probes generated from two independent LPC preparations from sections on two slides of the same biopsy (no. 2068). After data analysis with AIDA matrix the pairwise comparison of the normalized expression data was performed by the AIDA compare module. The detection threshold was set to 16% and 15% for no. 2068-1 and 2068-2, respectively. The green and red line represents 3-fold downregulation or upregulation, respectively. Journal of Investigative Dermatology 2004 122, 361-368DOI: (10.1046/j.0022-202X.2004.22240.x) Copyright © 2004 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 2 Numbers of genes regulated during progression of melanocytic lesions clustered according to functional groups. Sorted according to the pattern of regulation: (1) induction from nevi towards melanoma (light gray); (2) induction from melanoma towards metastases (gray); (3) induction from nevi towards melanoma and further upregulation towards metastases (dark gray); (4) genes upregulated from nevi towards melanoma and subsequently suppressed in metastases (black). The groups contain the following gene functions: Metabolism, signaling/cell cycle, receptors/attachment, others, unknown function. Journal of Investigative Dermatology 2004 122, 361-368DOI: (10.1046/j.0022-202X.2004.22240.x) Copyright © 2004 The Society for Investigative Dermatology, Inc Terms and Conditions

Figure 3 Statistical clustering by canonical discriminance analysis. The first and second function discriminance values for each case are plotted. Nevi: red spots; melanoma: green triangles; metastasis: blue squares. The center of each cluster is marked by an open square in the appropriate color. Journal of Investigative Dermatology 2004 122, 361-368DOI: (10.1046/j.0022-202X.2004.22240.x) Copyright © 2004 The Society for Investigative Dermatology, Inc Terms and Conditions