Astrocyte and Macrophage Regulation of YKL-40 Expression and Cellular Response in Neuroinflammation
Version of Record online: 22 DEC 2011
© 2011 The Authors; Brain Pathology © 2011 International Society of Neuropathology
Volume 22, Issue 4, pages 530–546, July 2012
How to Cite
Bonneh-Barkay, D., Bissel, S. J., Kofler, J., Starkey, A., Wang, G. and Wiley, C. A. (2012), Astrocyte and Macrophage Regulation of YKL-40 Expression and Cellular Response in Neuroinflammation. Brain Pathology, 22: 530–546. doi: 10.1111/j.1750-3639.2011.00550.x
- Issue online: 14 JUN 2012
- Version of Record online: 22 DEC 2011
- Accepted manuscript online: 11 NOV 2011 06:20AM EST
- Received 21 June 2011; Accepted 21 October 2011; Published Online Article Accepted 11 November 2011
Figure S1. Validation of YKL-40 staining procedure. Macrophages were gated based on forward scatter (FS) and side scatter (SS) log parameters as shown in the left graph. The arrow points to analysis of macrophages that were stained using different protocols. The histogram shows YKL-40 staining (black-dashed line) of macrophages, which is easily distinguishable from the following control stains: YKL-40 is not detected without intracellular staining (ICS) permeablization of the cell (dark gray solid line). Unstained, permeabilized macrophages are similar to the other controls (filled dark gray peak). Staining of permeabilized macrophages with an irrelevant biotin-conjugated antibody (filled black peak) (A). Histogram shows the distinction of macrophages that were: not treated with GolgiStop and stained with YKL-40 (filled light gray peak), stained with anti-biotin antibody only (filled dark gray peak), stained with 0.5 µg anti-YKL-40 biotin-conjugated antibody (dashed line) and stained with 5 µg anti-YKL-40 biotin-conjugated antibody (black line) (B). Expression of YKL-40 was not observed in CD3+ lymphocytes (C).
Figure S2. Validation of TNFα secretion in cocultured macrophages. Astrocytes were plated in chamber slides and were grown for 2 days before macrophages were added to the culture. Cocultured cells were grown for 14 days and the culture medium was collected every 2 days and analyzed for TNFα. Time course of TNFα levels released into the culture medium was determined by TNFα ELISA kit. Data shown are from a single experiment using duplicate samples and are representative of three independent experiments.
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