Metastasis compared to Primary Melanoma.

*<p>Proteins with significant changes greater than 20%.</p

LC/MS-based Label-free Protein Quantification Method.

*<p>Absolute Fold Change.</p>**<p>replicate.</p>***<p>replicate+sample.</p

Representative IHC staining of significant proteins found in melanomas.

<p>Anti-PKM1/M2 (rabbit) Ab: A) primary and B) brain metastasis. Anti-HSP90 (rabbit) Ab: C) primary and D) brain metastasis. Anti-MIF (mouse) Ab: E) primary and F) brain metastasis. Anti-STAT1 (mouse) Ab: G) primary and H) brain metastasis. I) Universal negative control rabbit Ig on primary melanoma tissue. J) Universal negative control mouse Ig on brain metastasis tissue. All magnifications were ×100. Scale bars represent 100 µm.</p

Representation of two ontological categories of differentially expressed proteins in metastasis by GeneGo pathways analysis.

<p>The results are ordered by −log10 of the p-value of the hypergeometric distribution. A) GeneGo canonical pathway maps, and B) GeneGo cellular process.</p

MS tracings of representative specific peptides.

<p>X-axis: retention time. Y-axis: peak intensity. A) Extracted ion chromatogram (XIC) for peptide GDYPLEAVR from pyruvate kinase isozymes M1/M2 of representative samples. Primary melanoma samples A.1 and A.2 are duplicate samples. Metastatic melanoma samples B.1 and B.2 are duplicates. Peak intensities were calculated by smoothing (as indicated by the green and red trace lines) and integrating the AUC of smoothed peaks for all samples within the same one-minute window (RT from 45.5 min to 46.5 min as indicated by the red trace line). B) Extracted ion chromatogram (XIC) for peptide ALLFVPR from heat shock protein HSP90AA2 of representative samples. Primary melanoma samples A.1 and A.2, and samples B.1 and B.2, are duplicate samples, respectively. Metastatic melanoma samples B.1 and B.2 are duplicates. RT from 59.7 min to 60.7 min as indicated by the red trace line.</p

Location and functional distribution of proteins isolated and identified from melanoma FFPE tissue.

<p>Proteins are categorized by A) cellular origin and, B) biological process, based on the Gene Ontology database.</p