(A, B) ... (A, B) The frequencies of Tregs from patients with SLE (n = 50) and healthy controls (n = 20) are shown. (C) Data plots show the correlation of Tregs with SLEDAI scores, serum anti-dsDNA and urine protein levels in patients with SLE. (D) Serum samples from SLE patients and healthy adults were assayed by ELISA for the levels of VCAM-1, ICAM-1 and VEGF. (E) The correlation of VCAM-1, ICAM-1 and VEGF with SLEDAI scores, serum anti-dsDNA and urine protein levels in patients with SLE are shown. (F) The relationship between the frequency of Tregs and the levels of soluble adhesion molecules and VEGF was analysed. Both correlation coefficient R and P values were calculated by the Pearson’s correlation test. Error bars represent the mean (s.e.m.). (n.s.: not significant, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001). Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) 2019. Published by Oxford University Press on behalf of the British Society for Rheumatology. All rights reserved. For permissions, please email: journals.permissions@oup.comThis article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model) Rheumatology (Oxford), kez112, https://doi.org/10.1093/rheumatology/kez112 The content of this slide may be subject to copyright: please see the slide notes for details.
Fig. 2 CD8+CD103+ iTregs show potent therapeutic effect on MRL/lpr lupus-prone nephritis ... CD8<sup>+</sup>CD103<sup>−</sup> med and CD8<sup>+</sup>CD103<sup>+</sup> iTregs induced from MRL/lpr mice were adoptively transferred to MRL/lpr mice at 6 and 8 weeks, respectively. (A) Measured body weight of ICR mice and MRL/lpr mice every 2 weeks. (B) Proteinuria was monitored by Uristix Test Strips. (C, D) The level of dsDNA Abs and total IgG titres in sera were measured by ELISA. (E) IF analysis of the IgG or C3 immune deposition in the glomeruli. IgG or C3 mean fluorescence intensity were shown. (F) Kidney pathology was analysed by haematoxylin and eosin, Masson, PAS, PAS-M and TEM. Representative pictures are shown. A one-way ANOVA comparison test was used for statistical analysis. There were six mice in each group. The data indicate the mean (s.e.m.) of three independent experiments. (n.s.: not significant, *P < 0.05, **P < 0.01, ****P < 0.0001). PAS: periodic acid–Schiff; PAS-M: periodic acid–Schiff methenamine; TEM: transmission electron microscopy; ANOVA: analysis of variance. Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) 2019. Published by Oxford University Press on behalf of the British Society for Rheumatology. All rights reserved. For permissions, please email: journals.permissions@oup.comThis article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model) Rheumatology (Oxford), kez112, https://doi.org/10.1093/rheumatology/kez112 The content of this slide may be subject to copyright: please see the slide notes for details.
Fig. 3 CD8+CD103+ iTregs treatment reduces the expression of pro-inflammatory mediator in the ... (A) qRT-PCR analysis of the mRNA levels of IL-17, TNF-α, IFN-γ, IL-6 and MCP-1 in the kidney. (B) Immunoblot analysis of the protein levels of IL-6 and MCP-1 in the kidney. (C) ELISA analysis of the expression levels of IL-17, TNF-α, IFN-γ, IL-6 and MCP-1 in the serum of lupus-prone mice. A one-way ANOVA comparison test was used for statistical analysis. There were six mice in each group. The data indicate mean (s.e.m.) of three independent experiments. (n.s. not significant, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001). ANOVA: analysis of variance. Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) 2019. Published by Oxford University Press on behalf of the British Society for Rheumatology. All rights reserved. For permissions, please email: journals.permissions@oup.comThis article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model) Rheumatology (Oxford), kez112, https://doi.org/10.1093/rheumatology/kez112 The content of this slide may be subject to copyright: please see the slide notes for details.
Fig. 4 CD8+CD103+ iTregs alleviate glomerular endothelial cell injuries in lupus-prone ... (A) qRT-PCR analysis of the mRNA levels of VCAM-1 and ICAM-1 in the kidney. (B) ELISA analysis of the expression levels of VCAM-1 and ICAM-1 in the serum of lupus-prone mice. (C) Immunoblot analysis of VCAM-1 and ICAM-1 protein levels in kidneys. (D) Images of kidney glomeruli from VCAM-1, ICAM-1 and CD31 IF. Red: VCAM-1, ICAM-1 and CD31, blue: DAPI. Scale bar: 100 µm. The capillary density in the glomeruli was analysed by the number of microvessels per cm<sup>2</sup>. (E) qRT-PCR and immunoblot analysis of VEGF levels in kidneys of mice. The levels of protein were normalized with β-actin. A one-way ANOVA comparison test was used for statistical analysis. There were six mice in each group. The data indicate the mean (s.e.m.) of three independent experiments. (n.s.: not significant, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001). ANOVA: analysis of variance. Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) 2019. Published by Oxford University Press on behalf of the British Society for Rheumatology. All rights reserved. For permissions, please email: journals.permissions@oup.comThis article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model) Rheumatology (Oxford), kez112, https://doi.org/10.1093/rheumatology/kez112 The content of this slide may be subject to copyright: please see the slide notes for details.
Fig. 5 Effects of CD8+CD103+ iTregs and CD8+CD103− meds on angiogenesis ... The proliferative effects of lupus-MGECs treated with the supernatant of CD8<sup>+</sup>CD103<sup>+</sup> iTregs and CD8<sup>+</sup>CD103<sup>−</sup> meds measured by EdU (A) and CCK8 absorbance (B). (C) The tube formation assay was used to analyse the vascular formation ability of lupus-MGECs. (D) The scratch wound assay was used to analyse endothelial cell migration and analysed by the ratio of non-migrated area divided by the baseline wound area. (E) The Transwell assay also revealed the same trend as the wound assay and was analysed via the number of migrated cells. Scale bar: 200 µm. (F) Western blot, qRT-PCR and ELISA were used to analyse the expression of VEGF. A one-way ANOVA comparison test was used for statistical analysis. The data indicate the mean (s.e.m.) of three independent experiments. (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001). ANOVA: analysis of variance; MGECs: mice glomerular endothelial cell. Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) 2019. Published by Oxford University Press on behalf of the British Society for Rheumatology. All rights reserved. For permissions, please email: journals.permissions@oup.comThis article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model (https://academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model) Rheumatology (Oxford), kez112, https://doi.org/10.1093/rheumatology/kez112 The content of this slide may be subject to copyright: please see the slide notes for details.