Semiquantitatively evaluated detection rate and sensitivity of IHC (anti-sstr2 and sstr5) and LDS using FITC-octreotate in tumor tissues.

<p>Semiquantitatively evaluated detection rate and sensitivity of IHC (anti-sstr2 and sstr5) and LDS using FITC-octreotate in tumor tissues.</p

Double stain analysis.

<p>Double stain analysis with FITC-octreotate and anti-sstr2a antibody. Immunofluorescence image of H69 cells on the slide were stained with DAPI (blue), FITC-octreotate (green) and Alexa 568-anti-sstrR2a antibody (red). The solid arrows indicate the double-stained cells with FITC-octreotate and anti-sstr2a antibody.</p

Synthesis of FITC-octreotate and evaluation of affinity.

<p>Synthetic route for FITC-octreotate. (a) FITC-octreotate was synthesized using the Fmoc solid phase method from ChemMatrix resin (CM) attached to the first amino acid. β-Ala and PEG2-Suc were introduced as linkers at the amino group after peptide chain elongation. FITC was modified at the N terminus. The protected peptide resin was treated with TFA. A disulfide bond was formed by the air oxidation method. (b) The RP-HPLC elution profile of FITC-octreotate. Column: Cosmosil 5C18 ARII (4.6 X 250mm) (Nacalai Tesque. Kyoto. Japan): eluent: 0.1% TFA in aqueous acetonitrile using the linear gradient indicated on the chromatogram flow rate: 1 mL/min. The arrow indicates FITC-octreotate. The product obtained was the isomer of FITC-octreotate caused by using the 5/6-FITC mixed isomer. (c) Saturation binding assay using FITC-octreotate on Somatostatin receptor subtype 2a. (d) Amido black stain and WLB membranes transferring sstr2 recombinant protein after SDS-PAGE.</p

Detection status of sstr in cases with neuroendocrine tumor.

<p>Detection status of sstr in cases with neuroendocrine tumor.</p

WB and WLB analysis.

<p>A Western blot analysis of the somatostatin receptor using anti-sstr2 (lane sstr2) and anti-sstr5 (lane sstr5) antibodies and a Western ligand blot analysis with FITC-octreotate (lane WLB) of a small-cell lung cancer cell (H69) lysate. All experiments were performed under the same conditions. The locations of the bands in the western blot analysis (the solid arrows of lane sstr2 and the dashed arrow of lane sstr5) correspond approximately to those in the western ligand blot analysis (the solid arrows and dashed arrow of lane WLB). The bands in the western ligand blot analysis (the solid arrows and dashed arrow of lane WLB) were completely blocked by the addition of a large amount of octreotide (X10000) (lane Blocking).</p

Comparison of IHC with LDS using FFPE sections.

<p>Comparison of immunohistochemistry using anti-sstr2 (b, f, j, n) and anti-sstr5 (c, g, k, o) antibodies with ligand derivative staining using FITC-octreotate (d, h, l, p) in myenteric plexuses, vascular endothelial cells, islets of the pancreas, and neuroendocrine tumors. Corresponding tissues were also stained by HE (a, e, i, m). All FFPE tissue sections were cut at a thickness of 3 μm from human pathological samples. The arrows (e, f, g, h) indicate myenteric plexuses and arrows (i, j, k, l) indicate vascular endothelial cells.</p

Additional file 1: of Splice-site mutation causing partial retention of intron in the FLCN gene in Birt-Hogg-Dubé syndrome: a case report

Figure S1. Amino acid sequence predicted by intron retention. Colored nucleotides are exons 9 and 10, and gray nucleotides starting from the mutated adenine (A, indicated by an arrow) are intron insertions. The predicted amino acid sequence is noted below codons in bold. A 130 bp intron retention leads to a frameshift from the beginning of exon 10, which results in premature termination (indicated by a rectangle). (PPTX 43 kb

List of proteomic analysis for the control liver in male group.

<p>Top 45 proteins exhibiting the probability of more than 0.8 are shown (5 family proteins are included). Proteins in bold letters were observed in the control liver in female group. Pre; precursor.</p><p>List of proteomic analysis for the control liver in male group.</p

Common down-regulated proteins in the liver and kidney after cadmium exposure.

<p>Proteins exhibiting the probability of more than 0.8 are shown. Pre; precursor.</p><p>Common down-regulated proteins in the liver and kidney after cadmium exposure.</p

Microscopic examination of the liver after oral chronic administration of cadmium.

<p>Left side; control group, right side; experimental group, upper side; female mice, and lower side; male mice. Scale bar is 50 μm.</p