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. 2016 Dec 1;7:552. doi: 10.3389/fimmu.2016.00552

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Copyright © 2016 Yang, Zhang, Qin, Wu, Li, Zhu, Ning, Lei and Shen.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Grp78-treated CD11c⁺ cells retained a regulatory signature after proinflammatory stimulation by LPS in vitro. LPS was added for the final 18–24 h during cell culture to generate DC_Grp78+LPS or DC_LPS. (A) Flow cytometry analysis of MHC-II, CD83, CD80, and CD40 expression on DC_Grp78+LPS and DC_LPS cells. Mean fluorescence intensity and (upper), representative flow cytometric dot plots (middle), and percentage of positive cells (lower) were depicted. (B) Percentage of B7-H3/B7-H4/B7-H1/B7-DC cells in DC_Grp78+LPS and DC_LPS cells. (C) Endocytosis assessed by 1 h incubation with dextran-rhodamine. MFI (left) and percentage of rhodamine positive cells (right) were depicted. (D) Cell culture supernatants were analyzed for IL-10, TGF-β, TNF-α, NO, HMGB-1, and IFN-γ by ELISA. (E) Gene expression of COX-2, TGF-β, TNF-α, iNOS, and ARG-1 assessed by qPCR. All results were representative of three independent experiments. Error bars were presented in SEM. *P < 0.05 vs. DC_LPS, **P < 0.01 vs. DC_LPS.

Grp78-treated CD11c⁺ cells retained a regulatory signature after proinflammatory stimulation by LPS in vitro. LPS was added for the final 18–24 h during cell culture to generate DC_Grp78+LPS or DC_LPS. (A) Flow cytometry analysis of MHC-II, CD83, CD80, and CD40 expression on DC_Grp78+LPS and DC_LPS cells. Mean fluorescence intensity and (upper), representative flow cytometric dot plots (middle), and percentage of positive cells (lower) were depicted. (B) Percentage of B7-H3/B7-H4/B7-H1/B7-DC cells in DC_Grp78+LPS and DC_LPS cells. (C) Endocytosis assessed by 1 h incubation with dextran-rhodamine. MFI (left) and percentage of rhodamine positive cells (right) were depicted. (D) Cell culture supernatants were analyzed for IL-10, TGF-β, TNF-α, NO, HMGB-1, and IFN-γ by ELISA. (E) Gene expression of COX-2, TGF-β, TNF-α, iNOS, and ARG-1 assessed by qPCR. All results were representative of three independent experiments. Error bars were presented in SEM. *P < 0.05 vs. DC_LPS, **P < 0.01 vs. DC_LPS.