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. 2014 Mar 13;9(3):e91705. doi: 10.1371/journal.pone.0091705

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© 2014 Weiterer et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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A: In RT-PCR experiments using whole tracheal preparations we found evidence for mRNA of TNF-R1 but not TNF-R2 (n = 3, bp: 100 kD). B: In tracheae previously exposed to TNF-α over 60 min we observed a non-significant increase in TNF-R1 mRNA when using qPCR (p = 0.2, n = 4). C: In addition immunoreaction against TNF-R1 was used to identify the anatomical location of the TNF-R1. Immunoreaction against TNF-R1 stained only cells within the epithelial layer. These were either found as isolated cells or as clusters of 2–4 cells (D). E: Pre-absorption of the primary antibody with TNF-R1 peptide did not stain any cells in the tracheal epithelium. F: In addition experiments omitting primary antibody were used as negative controls evoked no immunoreactive staining of epithelial structures. (Scale bars: 20 μm).