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BioMed Research International
Volume 2013 (2013), Article ID 654354, 11 pages
http://dx.doi.org/10.1155/2013/654354
Research Article

Increased Expression of Epidermal Growth Factor Receptor (EGF-R) in Patients with Different Forms of Lung Fibrosis

1Department of Pneumonology, University Hospital of Alexandroupolis, Medical school, Democritus University of Thrace, 68100 Alexandroupolis, Greece
2Department of Pathology, Veterans Administration Hospital (NIMTS), 11521 Athens, Greece
3Department of Cardiothoracic Surgery, University Hospital of Alexandroupolis, Medical School, Democritus University of Thrace, 68100 Alexandroupolis, Greece
4Department of Pathology, University Hospital of Alexandroupolis, Medical school, Democritus University of Thrace, 68100 Alexandroupolis, Greece

Received 18 April 2013; Accepted 21 May 2013

Academic Editor: Shoichiro Ono

Copyright © 2013 Argyris Tzouvelekis et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Supplementary Material

Methods: Tissue Microarrays: Tissue samples were snap frozen and stored at 70° C. Specimens were fixed in cold-ethanol for 16 h and then embedded in paraffin. Tissue cylinders of 1.5 mm diameter were punched from selected areas of each “donor” block by utilizing a thin-wall stainless tube from a precision instrument (TMA-100, Chemicon, USA (29). Ultimately, we created two tissue microarray blocks comprising of 100 tissue elements each. Immunohistochemistry: The slides were deparaffinised and En Vision immunohistochemistry protocol (DAKI corp, Denmark) was carried through the use of an automated immunohistochemistry staining system (Bond-Biogenex, USA), as previously described (30-32). Briefly, this immunohistochemistry protocol is based on a water-soluble, dextran polymer system preventing the endogenous biotin reaction, which is responsible for the background in the stained slides. Quantitative Real-Time reverse transcriptase-polymerase chain reaction (qRT-PCR): RNA expression values (also referred to as relative mRNA expression) were calculated as ratios (differences between the Ct values) between Egfr and COL1A2 and B2M and b-actin, respectively, that served as housekeeping genes.

  1. Supplementary Material