Artesunate Restrains Maturation of Dendritic Cells and Ameliorates Heart Transplantation‐Induced Acute Rejection in Mice through the PERK/ATF4/CHOP Signaling Pathway

Figures

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  Figure 1. (a) Chemical molecule structure diagram of ART; (b) survival curve of grafts (n = 6); (c) the appearance of cervical lymph nodes and spleen in different groups; (d) weight of spleens from different groups; (e) HE staining of heart tissue and statistics graph of cells' number per unit (∗P < 0.05 compared with NC; #P < 0.05 compared with HT).

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  Figure 2. (a) Immunofluorescence staining and mean fluorescence intensity of CD4 in grafts (blue: DAPI, red: CD4); (b) immunofluorescence staining and mean fluorescence intensity of CD8 in grafts (blue: DAPI, red: CD8); (c) immunohistochemistry of FOXP3 and statistics graph of positive cells' number per unit (∗P < 0.05 compared with NC; #P < 0.05 compared with HT).

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  Figure 3. (a) Flow cytometric sorting of CD4+/8+ T cells in spleens and statistics chart; (b) flow cytometric sorting of CD4+/8+ T cells in LNs and statistics chart; (c) CD4+CD25+FOXP3+ cells in spleens and statistics chart; (d) CD4+CD25+FOXP3+ cells in LNs and statistics chart (∗P < 0.05 compared with NC; #P < 0.05 compared with HT).

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  Figure 4. (a) TUNEL staining and mean fluorescence intensity of the heart (blue: DAPI, red: TUNEL); (b) 8-OHdg staining mean fluorescence intensity of the heart (blue: DAPI, red: 8-OHdg; ∗P < 0.05 compared with NC; #P < 0.05 compared with HT); (c) ROS of DCs after stimulation with LPS (∗∗P < 0.01 compared with CON; ##P < 0.01 compared with LPS).

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  Figure 5. (a) Flow cytometric sorting of CD11c+ and CD86+ cells in lymph nodes; (b) flow cytometric sorting of CD11c+ and MHC-II+ cells in lymph nodes (∗P < 0.05 compared with NC; #P < 0.05 compared with HT); (c) flow cytometric sorting of CD11c+ and CD86+ cells in BMDCs; (d) flow cytometric sorting of CD11c+ and MHC-II+ cells in BMDCs (∗P < 0.05 compared with CON; #P < 0.05 compared with LPS); (e) immunofluorescence staining and mean fluorescence intensity of MHC-II in BMDCs (blue: DAPI, red: MHC-II); (f) western blotting of PERK/ATF4/CHOP signaling pathway (#P < 0.05 compared with LPS).

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  Figure 6. (a) ROS of DCs after LPS and CCT (∗∗P < 0.01 compared with CON; ##P < 0.01 compared with LPS); (b) flow cytometric sorting of CD11c+ and CD86+ cells in BMDCs; (c) flow cytometric sorting of CD11c+ and MHC-II+ cells in BMDCs (∗P < 0.05 compared with CON; #P < 0.05 compared with LPS); (d) immunofluorescence staining and mean fluorescence intensity of MHC-II in BMDCs (blue: DAPI, red: MHC-II).

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  Figure 7. The mechanism of ART in BMDCs. ART reduces the generation of ROS to alleviate the endoplasmic reticulum stress and inhibits the maturation of DCs via reducing the expression of the PERK/ATF4/CHOP protein.