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Blood, Vol. 107, Issue 3, 1010-1017, February 1, 2006
A novel form of immune signaling revealed by transmission of the inflammatory mediator serotonin between dendritic cells and T cells
Blood O'Connell et al.
107: 1010
Supplemental materials for: O’Connell et al, Vol 107, No 3, 1010-1017
Files in this Data Supplement:
- Figure S1. Nonspecific labeling controls for SERT expression by BMDCs (JPG, 92 KB)
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Density gradient-enriched d5 BMDCs were immunolabeled and analyzed by confocal microscopy. Fluorescent micrographs are overlays of red, green, and blue channels to illustrate specific versus nonspecific labeling. The upper left panel shows CD11c+ (hamster anti-CD11c + biotinylated rabbit anti-hamster IgG + SA-Alexa Fluor 488) and SERT— control (mouse IgG2b + Xenon Alexa Fluor 546). The upper right panel shows CD11c— control (hamster IgG1 + biotinylated rabbit anti-hamster IgG + SA-Alexa Fluor 488) and SERT+ control (rabbit anti-SERT + Xenon Alexa Fluor 546). The lower left panel shows CD11c— control (hamster IgG1 + biotinylated rabbit anti-hamster IgG + SA-Alexa Fluor 488), SERT— control (mouse IgG2b + Xenon Alexa Fluor 546) and nuclear counterstain (TO-PRO-3; blue). In the lower right panel, the bright field image is overlaid with the nuclear counterstain (TO-PRO-3). Scale bar represents 20 µm.
- Figure S2. Nonspecific labeling controls for 5HT uptake by BMDCs (JPG, 60 KB)
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Day-5 BMDCs were purified by FACS sorting for CD11c-PE-Cy7+ cells and were then loaded with 5-HT (100 µM). Fluorescent micrographs are overlays of red, green, and blue channels to illustrate specific versus nonspecific labelling. The upper left panel shows LAMP-1+ control (rat anti-LAMP-1 + goat anti-rat IgG Alexa Fluor 488) and 5HT— control (5HT-absorbed rabbit anti-5HT + biotinylated goat anti-rabbit IgG + SA-Alexa Fluor 546). The upper right panel shows LAMP-1+ control (rat IgG2a + goat anti-rat IgG Alexa Fluor 488) and 5HT+ control (rabbit anti-5HT + biotinylated goat anti-rabbit IgG + SA-Alexa Fluor 546). The lower left panel shows LAMP-1— control (rat IgG2a + goat anti-rat IgG Alexa Fluor 488), 5HT— control (5HT-absorbed rabbit anti-5HT + biotinylated goat anti-rabbit IgG + SA-Alexa Fluor 546) and nuclear counterstain (TO-PRO-3; blue). In the lower right panel, the bright field image is overlaid with the nuclear counterstain (TO-PRO-3). Scale bar represents 10 µm.
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