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. 2020 May 5;11(5):319. doi: 10.1038/s41419-020-2547-4

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© The Author(s) 2020

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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Fig. 5 — a JC-1 staining was used to detect mitochondrial membrane potential of MPCs exposed to LN plasma. Scale bars: 10 μm. b Quantification of JC-1 fluorescence (red:green ratio) showing changes in fluorescence intensity in MPCs exposed to LN plasma in different time. JC-1 fluorescence was normalized with the red-to-green ratio of 0 h group. *P < 0.05 vs. 0 h group, ^#P < 0.05 vs. 12 h group, ^△P < 0.05 vs. 48 h group (n = 20). c ATP assay showed the ATP content in MPCs exposed to LN plasma. *P < 0.05 vs. 12 h group, ^#P < 0.05 vs. 48 h group (n = 3). d, e Western blot assay showed the expression of Drp1 and Mfn1 in the MPCs exposed to LN plasma. *P < 0.05 vs. 0 h group (n = 3). f–h IHC staining showed the expression of Drp1 and Mfn1 in LN patients. Scale bars: 50 μm. *P < 0.05 vs. control group. i Mito Tracker staining showed the mitochondria morphology of MPCs after nestin overexpression and downregulation and exposed to LN plasma for 24 h. Scale bars: 10 μm. j Quantification of Mito Tracker staining (aspect ratio) in MPCs in different treatment. *P < 0.05 vs. control plasma group, ^#P < 0.05 vs. LN plasma + shNC group, ^△P < 0.05 vs. LN plasma + NC group (n = 360). k Quantification of Mito Tracker staining showing the different morphology mitochondrial numbers in MPCs in different treatment. *P < 0.05 vs. control plasma group, ^#P < 0.05 vs. LN plasma + shNC group, ^△P < 0.05 vs. LN plasma + NC group (n = 20). l JC-1 staining was used to detect mitochondrial membrane potential of MPCs after downregulation of the nestin expression and exposed to LN plasma for 24 h. Scale bars: 10 μm. m Quantification of JC-1 fluorescence (red:green ratio) showing changes in fluorescence intensity in MPCs in different treatment. JC-1 fluorescence was normalized with the red-to-green ratio of the control plasma group. #P < 0.05 vs. LN plasma + shNC group (n = 20). n, o Western blot assay showed the downregulation of nestin affected the expression of Drp1 and Mfn1 in the MPCs exposed to LN plasma for 24 h. *P < 0.05 vs. control plasma group, ^#P < 0.05 vs. LN plasma + shNC group (n = 3). p, q Western blot assay showed the downregulation of nestin affected the expression of Drp1 and Mfn1 in the MPCs. *P < 0.05 vs. control FBS group (n = 3). Student’s t test and Mann−Whitney nonparametric tests were applied to compare variables between two groups. Bonferroni’s correction was performed to analyze statistical significance between multiple comparisons.