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. 2019 Sep 25;9:13825. doi: 10.1038/s41598-019-50311-z

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Long-term culture in SMG reduces the T cell stimulatory capacity of JAWS II DC. (a) JAWS II cells (2 × 10⁵/ml) were cultured in Static (light gray) or SMG (dark gray) conditions for 5 days. DC were collected and stained with antibodies for MHC class I and II, CD80 and CD86 and their corresponding isotype controls. Open histograms with broken lines represent isotype controls. (b) Graph represents the MFI of each of the surface molecules examined in (a) for Static (light gray bars) and SMG JAWS II DC (black bars). (c) JAWS II DC (2 × 10⁵/ml) were cultured in Static (light gray bars) or SMG (black bars) conditions for 7, 12 and 14 days. At each time point, Static and SMG JAWS II DC were harvested, washed and incubated with OT-II CD4⁺ TCH and OVA323-339 for an additional 24 hours in Static conditions. Culture supernatants were collected and assessed for IL-2 production by ELISA. One representative experiment of two independent experiments with similar results is shown for (a). The data in (b) represents the mean + SD of quadruplicates of two independent experiments. In (b), *p-value ≤ 0.05 comparing the expression of surface molecules by Static and SMG JAWS II DC. In (c), *p-value ≤ 0.05 comparing the activation of T cell IL-2 production by day 12 SMG and day 14 SMG JAWS II DC as well as day 14 Static and SMG JAWS II DC.