Patient demographics, resection location and tumor types for the eight NSCLC analyzed samples.

<p>Patient demographics, resection location and tumor types for the eight NSCLC analyzed samples.</p

Relative changes in protein expression for 820 proteins from eight NSCLC resection samples.

<p>Signal differences between adjacent and distant tissue (panel A), tumor and adjacent tissue (panel B) and tumor and distant tissue (panel C) are expressed as log₂ median ratios. The dotted line represents two-fold change (log₂ = 1).</p

Categorization of NSCLC tissue biomarkers into biological major processes.

*<p>Novel NSCLC Biomarker.</p

SOMAmer histochemistry on frozen tissue sections for selected biomarkers detected in this study.

<p>(A) Thrombospondin-2 SOMAmer (red) staining the fibrocollagenous matrix surrounding a tumor nest. (B) Corresponding normal lung specimen stained with Thrombospondin-2 SOMAmer (red). (C) Macrophage mannose receptor SOMAmer (red) staining scattered macrophages in a lung adenocarcinoma. (D) Macrophage Mannose Receptor SOMAmer (red) staining numerous alveolar macrophages in a section of normal lung parenchyma. (E) Multicolor image highlighting the cytomorphologic distribution of macrophage mannose receptor SOMAmer staining: Green = Cytokeratin (AE1/AE3 antibody), Red = CD31 (EP3095 Antibody), and Orange = SOMAmer. All nuclei in this figure are counterstained with DAPI.</p

Box plots of SOMAmer signals in the tissue homogenates.

<p>Proteins with increased (panel A) or decreased (panel B) levels in tumor tissue compared with adjacent or distal tissue (panel A) from eight NSCLC samples used in this study. Each individual is indicated with a different symbol. The horizontal lines of each box correspond to the first, second, and third quartiles (25%/50%/75%) and the whiskers correspond to the maximum and minimum values.</p

Thrombospondin-2 (TSP-2) histochemical identification in tissue samples.

<p>TSP-2 is identified in serial frozen sections of a single lung carcinoma specimen by (A) a home-made rabbit polyclonal TSP-2 polyclonal antibody, (B) the pre-immune serum from rabbits used to make the home-made polyclonal antibody, (C) a commercial (Novus) rabbit polyclonal TSP-2 antibody, and (D) the TSP-2 SOMAmer. The TSP-2 SOMAmer was used to stain frozen sections of normal and malignant lung tissue, with standard Avidin-Biotin-Peroxidase color development, to demonstrate different morphologic distributions: (E) Strong staining of the fibrotic stroma surrounding tumor nests, with minimal cytosolic staining of carcinoma cells, (F) Strong staining of the fibrotic stroma surrounding tumor nests in a mucinous adenocarcinoma, with no significant staining of the carcinoma cells, (G) normal lung tissue, showing strong cytosolic staining of bronchial epithelium and scattered alveolar macrophages, and (H) strong cytosolic staining of an adenocarcinoma, with no significant staining of the non-fibrotic, predominantly inflammatory stroma.</p

Plot of the cumulative density function (CDF) for the coefficient of variation (CV) between triplicate samples.

<p>The tumor, adjacent non-tumor, and distant non-tumor tissue resections were sampled, extracted, and analyzed with the SOMAscan proteomic assay in triplicate for two individuals in the study. The median CV for all 6 triplicates was 4.5% (black line).</p

List of potential NSCLC biomarkers identified from serum and tissue samples.

<p>Proteins identified in both studies are shown in boldface font.</p

Changes in protein expression in NSCLC tissue compared to serum.

<p>The top two panels show the log2 ratio (LR) derived from serum samples versus log ratios derived from adjacent tissue and distant tissue, respectively. The bottom four panels feature zoomed portions of plots above, indicated by the color of the plot (green for decreased and red for increased expression compared to non-tumor tissue). Analytes shown in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0035157#pone-0035157-g002" target="_blank">Figure 2</a> have been labeled and analytes mentioned in the publication on the serum samples are shown in filled red symbols red.</p

ESAM histochemical staining in tissue samples.

<p>ESAM staining is shown in lung tumor (A,C) and normal lung (B,D) distant from the tumor. Endothelial cells are visibly more abundant in the normal lung section, consistent with the high vascularity of normal lung. Raw images are shown in A and C, with ESAM-positive cells identified by the CellProfiler algorithm marked with a “1” in images B and D.</p