Figure 3 | Scientific Reports

Figure 3

From: Improved Detection of Cytokines Produced by Invariant NKT Cells

Figure 3

Effective detection of iNKT cell IL-10 required the removal of dead cells. (A) C57BL/6 splenocytes and PBMCs were stimulated in vitro for 4 h with PMA and ionomycin and the percentage of IL-10+ iNKT cells was measured by ICCS. (B) C57BL/6 splenocytes were either left untreated (control) or purified via a density-gradient (+Lymphoprep) and stimulated in vitro for 4 h with PMA and ionomycin in the presence of the Golgi-transport inhibitors Brefeldin A and monensin. The percentage of IL-10+ iNKT cells was then measured by ICCS. (C) C57BL/6 animals were injected i.v. with 1μg αGalCer and 90 min later splenocytes were purified via a density-gradient and incubation for 2 h in vitro in the presence of the Golgi-transport inhibitors Brefeldin A and monensin. The percentage of IL-10+ iNKT cells was then measured by ICCS. A summary graph (left panel) and representative data (right panel) are shown, respectively. The means of activation is also given on the left of the panels. Representative data from one of at least three independent experiments are shown.

Back to article page