Additional corresponding author: Dr. Miguel A. Lasunción, E-mail: firstname.lastname@example.org
Curcumin promotes exosomes/microvesicles secretion that attenuates lysosomal cholesterol traffic impairment
Version of Record online: 29 NOV 2013
© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Molecular Nutrition & Food Research
Volume 58, Issue 4, pages 687–697, April 2014
How to Cite
Canfrán-Duque, A., Pastor, Ó., Quintana-Portillo, R., Lerma, M., de la Peña, G., Martín-Hidalgo, A., Fernández-Hernando, C., Lasunción, M. A. and Busto, R. (2014), Curcumin promotes exosomes/microvesicles secretion that attenuates lysosomal cholesterol traffic impairment. Mol. Nutr. Food Res., 58: 687–697. doi: 10.1002/mnfr.201300350
- Issue online: 1 APR 2014
- Version of Record online: 29 NOV 2013
- Manuscript Accepted: 20 SEP 2013
- Manuscript Revised: 10 SEP 2013
- Manuscript Received: 13 MAY 2013
- Ministerio de Ciencia e Innovación, Spain. Grant Number: SAF2011-29951 and SAF2009-08764
- National Institutes of Health. Grant Number: R01HL107953 and R01HL106063
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Figure S1. Effects of the drugs used on cell viability HepG2 and THP-1 macrophages cells. Cells were exposed to LDL (30 μg/ml of cholesterol) in the absence (control) or the presence of 5 μM U18666A and 5% DMSO for a total 24-h period. Where indicated, during the last 2 or 1 h, the cells were treated with 30 μM curcumin or 1 μM thapsigargin, respectively. Then, the mitochondrial activity (XTT assay) was measured after the treatment. Values are the mean ± S.E.M of three independent experiments with three replicates each. Statistical comparisons are shown versus control (* P < 0.05).
Figure S2. Effects of curcumin on the intracellular distribution of LDL in THP-1 differentiated macrophages cells treated with U18666A. The cells were exposed to DiI-LDL (30 μg/ml of cholesterol) for 24 h in the absence (control) or the presence of 5 μM U18666A for a total 24-h period. Where indicated, during the last 2 h, cells were treated with 30 μM curcumin. Then, the cells were washed, fixed and stained with filipin and CD63 and analyzed using a confocal microscope.
Figure S4. Analysis of the purity of exosome/microvesicles preparations. THP-1 differentiated macrophages were exposed to LDL (30 μg/ml of cholesterol) in the absence (control) or the presence of 5 μM U18666A for 22 h. Then the medium was removed, cells were washed twice, and serum-free medium was added, supplemented or not (control) with U18666A, and incubation was continued for 2 h in the presence of 30 μM curcumin (Cur) or 1 μM thapsigargin (Tg) (last 1 h), as indicated. At the end of the incubation, the medium was removed, and the exosomes/microvesicles were isolated and calnexin, a reticulum endoplasmic marker, and flotillin-2 were analyzed by western blot. In the same gel, control cell lysate was also analyzed for comparison. Results from a representative experiment of 3 independent experiments are shown.
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