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Mediators of Inflammation
Volume 2016, Article ID 9858374, 12 pages
Research Article

Regulation of IL-33 by Oncostatin M in Mouse Lung Epithelial Cells

McMaster Immunology Research Centre, Department of Pathology and Molecular Medicine, McMaster University, Hamilton, ON, Canada

Received 21 June 2016; Revised 12 August 2016; Accepted 17 August 2016

Academic Editor: Alex Kleinjan

Copyright © 2016 Carl D. Richards 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

Supplemental Figure 1 demonstrates an induction of full-length IL-33 protein (35 kDa) and phospho-STAT3 (79, 86 kDa) by Western blot, in whole lung homogenates of C57Bl/6 mice treated with AdOSM, in comparison to AdDel70 control vector. Samples were also probed for β-Actin and total-STAT3 as loading controls.

Supplemental Figure 2 shows that the antibody in Western blots detected prominent bands corresponding to full length IL-33 (approximately 35kDa) induced by OSM whereas mature IL-33 signal (recombinant mature IL-33 at approximately 18-19 kDa) was absent/low in mouse lung extracts of both AdDel70 and AdOSM-treated BALB/c mice, as well as C10 cell culture lysates stimulated with OSM. Recombinant mature IL-33 (R&D) was run on the same gel for comparison and samples were probed for β-Actin as a loading control.

Supplemental Figure 3 contains results from IHC staining for IL-33 in C10 cell cultures in vitro. The data showed detectable IL-33 in nuclei of un-stimulated cells and a marked increase in nuclear staining upon OSM stimulation.

Supplemental Figure 4 confirms that AdOSM treatment resulted in the same effects in BALB/c mouse lung as previously described in this system [31], including inflammatory cell accumulation in broncho-alveolar lavage (BAL) fluid and histopathology of lung tissue sections. The results indicated expected elevation of cells in the BAL fluid as well as increases in epithelial cell hyperplasia, thicker alveolar walls, and inflammatory cell accumulation in tissue sections, in response to AdOSM.

  1. Supplementary Figures