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. 2016 Jan 19;15(4):1246–1261. doi: 10.1074/mcp.M115.054593

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© 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

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Fig. 3. — SEC-MAP identified 47 antigens that were differentially expressed in diverse subtypes of primary childhood acute leukemia samples. Forty-five proteins (including phosphorylated forms of CD45 and lymphocyte cytosolic protein 2, LCP2), and one glycolipid carbohydrate (stage-specific embryonic antigen-4, SSEA4) were differentially expressed in the three subtypes of primary AL (BCP-ALL, n = 35, T-ALL, n = 9, and AML, n = 13) according to SEC-MAP (p < 0.05). The heat map shows these 46 antigens (in rows, expressed as the sum of the MFI from PE-labeled streptavidin in the SEC fractions determining the entity) as measured by the SEC-MAP in 57 patient samples (columns). Hierarchical clustering with Euclidean distance metrics and average linkage was used for the analysis (A). Four markers were found to be included in adhesion and migration, 3 in proliferation, 1 in differentiation, 3 in apoptosis, 1 in ubiquitylation, 12 in cell signaling, 7 in transcription, and 1 in metabolism. Eleven CD antigens and two adapter molecules were identified (B). Drebrin (DBN1) was found to be highly expressed in BCP-ALL in comparison with AML (top panel). Glutamate dehydrogenase 1, mitochondrial (GLUD1) was found to be highly expressed in AML in comparison with BCP-ALL (middle panel). STAT5A was found to be highly expressed in T-ALL in comparison with BCP-ALL (bottom panel). The line plots show two representatives of BCP-ALL (solid line) and AML (dashed) (top panel), AML (solid line) and BCP-ALL (dashed) (middle panel) and T-ALL (solid line) and BCP-ALL (dashed) (bottom panel). The x axis represents 24 SEC fractions, and the y axis indicates the MFI of PE-labeled streptavidin. The protein entities of DBN1, GLUD1 and STAT5A are marked by vertical lines. The boxplots show the expression of DBN1, GLUD1 and STAT5A in the entire cohort of examined patient samples (BCP-ALL, n = 35; T-ALL, n = 9; and AML, n = 13) (C). Outcome Predictor in Acute Leukemia 1 (OPAL1 or WBP1L) was identified as highly expressed in ETV6-RUNX1-positive BCP-ALL samples (n = 6) in comparison with ETV6-RUNX1-negative BCP-ALL samples (n = 29) at the protein level (p < 0.05). The line plot shows the OPAL1 expression as measured by SEC-MAP in two representative ETV6-RUNX1-positive BCP-ALL samples (solid line) and two representative ETV6-RUNX1 negative BCP-ALL samples (dashed). The x axis represents 24 SEC fractions, and the y axis indicates the MFI of PE-labeled streptavidin. The protein entity of OPAL1 is marked by vertical lines. The boxplot shows the OPAL1 expression in the whole cohort of BCP-ALL samples (D).