Adipocyte-derived lipids modulate CD4+ T-cell function
Version of Record online: 24 APR 2013
© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
European Journal of Immunology
Volume 43, Issue 6, pages 1578–1587, June 2013
How to Cite
Ioan-Facsinay, A., Kwekkeboom, J. C., Westhoff, S., Giera, M., Rombouts, Y., van Harmelen, V., Huizinga, T. W. J., Deelder, A., Kloppenburg, M. and Toes, R. E. M. (2013), Adipocyte-derived lipids modulate CD4+ T-cell function. Eur. J. Immunol., 43: 1578–1587. doi: 10.1002/eji.201243096
- Issue online: 21 JUN 2013
- Version of Record online: 24 APR 2013
- Accepted manuscript online: 15 MAR 2013 03:30AM EST
- Manuscript Accepted: 8 MAR 2013
- Manuscript Revised: 8 FEB 2013
- Manuscript Received: 26 OCT 2012
- The Netherlands Organisation for Scientific Research. Grant Number: 115142-2
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Fig. S1 A) Gating strategy for Fig.1A. A live gate based on FSC-A/SSC-A was first set on stromal vascular fraction cells (SVF), followed by two gates to exclude double cells. The intersection of this gate was used to exclude cells positive for dead cell discriminator, then this gate was used for gating the lymphocyte population based on FSC-A/SSC-A. Lymphocytes positive for CD3 and negative for CD14 were selected and, within this population, the CD4+CD8- cells were further analyzed as depicted in Fig. 11A. B) Gating strategy for figure 2B. A live gate based on cell FSC-A/SSC-A was set, followed by two gates to exclude double cells. Within the intersection of these gates, cells positive for dead cell discriminator were excluded as described in “Materials and Methods”. Cells in this live gate were further analyzed as depicted in Fig. 2B.
Fig. S2 Isolated CD4+ T cells were activated overnight with 5 μg/ml plate-bound anti-CD3 and 1 μg/ml soluble anti-CD28 in the presence or absence of ACM obtained from infrapatellar fat pad-derived adipocytes. Mean levels with SEM (n = 10) are depicted for IL-17 (A) and IL-5 (B) in cultures of activated T cells alone (T) or in the presence of ACM, corrected for the respective cytokines present in ACM (T+ACM corr.); p-values were calculated using Wilcoxon signed rank test.
Fig. S3 A) Chromatograms in positive ionization mode of lipids in the total lipid fraction of ACM or control medium (ctr). Arrows indicate differences between ACM and control medium and the signal corresponding to triglycerides. B) Glycerol was measured as an index of lipolysis induced by isoprenaline, noradrenaline (NA) or NA in combination with yohimbine (Yoh), as described in Materials and Methods. The response to the (anti)lipolytic agents was expressed as percentage of basal glycerol release. The average with SEM glycerol release of three different adipose tissues is presented. The curve represent the non-linear fit of the response. R2 = 0,89 for isoprenaline, R2 = 0,53 for NA.
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