Figure 1.
Diagram of experimental methods used for multiplexed barcoded antibody array.
The three cell lines were labeled with or without intracellular dye prior to admixing the cells into a single pool. The cells were then aliquoted into each well for antibody labeling. The contents of each well were then processed on a flow cytometer. The identity of each cell line was determined based on fluorescence intensity. The appropriate gates were drawn allowing for simultaneous analysis for each antibody. Histograms for mouse IgM isotype control are shown.
Table 1.
Broadly expressed tumor antigens.
Figure 2.
Oncomine heatmap analysis in 4 published datasets for expression of tumor antigens described in Table 1. Only those genes that were consistently upregulated across datasets with p<0.05 are shown. The numbers in parentheses indicate the number of samples analyzed. Abbreviations: normal colon, NC; ascending colon, AC; descending colon, DC; sigmoid colon, SC; transverse colon, TC (n = 1).
Figure 3.
Validation of integrin α6 expression in colon cancer by immunohistochemistry.
A) H&E (top left) and integrin α6 IHC (top right) from clinical colon cancer specimens at low magnification. Areas of normal mucosa (N) and adjacent primary colon cancer (P) are indicated. Lower panels provide higher magnification fields of integrin α6 in normal (left) and tumor (right). B) Representative examples of liver and lymph node metastases. The regions of colon cancer metastases (M) are visible by H&E (left) and corresponding staining with integrin α6 (right). An area of normal liver (L) is indicated. All lymph node samples contained a high degree of fibrosis around the lesion that displaced normal lymphoid tissue from the field of view. Scale bar: 50 µm.
Figure 4.
CD10 expression in SW480 versus SW620.
Histogram plots from antibody array for the CD10 antigen in SW480 (A) and SW620 (B). Red indicates isotype control while the blue line is staining for CD10. The number in the top left is the cell positivity. There is a clear shift from a small shoulder population in SW480 to complete binding in SW620 cells. C) Immunoblotting for CD10 confirms the strong change in CD10 expression.
Table 2.
Antigens upregulated in metastasis.
Table 3.
Antigens downregulated in metastasis.
Table 4.
Expression of surface stem cell markers.