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(A) Western ... (A) Western blot showing DTX1 knockdown with siRNA in human CD4<sup>+</sup> T cells. Actin was used as the loading control. (B) Purified.

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Presentation on theme: "(A) Western ... (A) Western blot showing DTX1 knockdown with siRNA in human CD4<sup>+</sup> T cells. Actin was used as the loading control. (B) Purified."— Presentation transcript:

1 (A) Western ... (A) Western blot showing DTX1 knockdown with siRNA in human CD4<sup>+</sup> T cells. Actin was used as the loading control. (B) Purified human CD4<sup>+</sup> T cells from peripheral blood were transfected with DTX1 or control (Ctrl) siRNA. Transfected T cells were stimulated by plate-bound anti-CD3 (0, 0.5, 1, 2, 4 μg/ml) and anti-CD28 antibodies (0, 0.5, 1, 2, 4 μg/ml) for 24 and 48 h. IL-2 and IFN-γ concentrations in supernatants were measured by ELISA. (C) Proliferation assay of stimulated (anti-CD3/28) CD4<sup>+</sup> T cells transfected by DTX1 and Ctrl siRNA. Each experiment was performed in triplicate. Experiments were independently repeated three times with similar results. *P < 0.05. Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) Published by Oxford University Press on behalf of the British Society for Rheumatology. All rights reserved. For permissions, please article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model ( Rheumatology (Oxford), Volume 58, Issue 4, 09 January 2019, Pages 719–728, The content of this slide may be subject to copyright: please see the slide notes for details.

2 (A) DTX1 ... (A) DTX1 expression in PBMCs from 100 SLE patients and 50 healthy controls was measured with quantitative RT-PCR and normalized to the GAPDH control (left). The ROC curve was analysed for SLE diagnosis using the DTX1 mRNA level (right). (B) The DTX1 level was significantly lower in SLE patients with severe active disease (SLEDAI-2K score >10) than in those with inactive disease (score <4) or mild/moderate disease activity (score 4–10). (C) Association between DTX1 level in PBMCs and disease activity in SLE patients (SLEDAI). Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) Published by Oxford University Press on behalf of the British Society for Rheumatology. All rights reserved. For permissions, please article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model ( Rheumatology (Oxford), Volume 58, Issue 4, 09 January 2019, Pages 719–728, The content of this slide may be subject to copyright: please see the slide notes for details.

3 The associations of ... The associations of DTX1 expression in PBMCs with clinical manifestations of SLE patients were investigated. (A) DTX1 expression was compared between 29 SLE patients with active LN and 16 SLE patients with inactive LN (left). The ROC curve was analysed for identification of activity status in SLE patients with LN using the DTX1 mRNA level (right). (B) DTX1 expression in SLE patients with or without lung involvement (left). The ROC curve was analysed for lung involvement diagnosis in SLE patients using the DTX1 mRNA level (right). (C) DTX1 gene expression was significantly lower in SLE patients with low complement C3 or C4 levels than in those with normal C3 or C4 levels. Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) Published by Oxford University Press on behalf of the British Society for Rheumatology. All rights reserved. For permissions, please article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model ( Rheumatology (Oxford), Volume 58, Issue 4, 09 January 2019, Pages 719–728, The content of this slide may be subject to copyright: please see the slide notes for details.

4 Fig. 4 IFN-γ production was increased in CD4+ T cells with low DTX1 expression in SLE ...
CD4<sup>+</sup> T cells were purified from PBMCs of six SLE patients and the DTX1 mRNA level was measured by quantitative RT-PCR. IL-2 and IFN-γ secretion by T cells was measured in SLE patients with high and low DTX1 mRNA levels in T cells. Purified human CD4<sup>+</sup> T cells with high and low DTX1 mRNA levels from SLE patients were stimulated by plate-bound anti-CD3 (0, 1, 2, 4 μg/ml) and anti-CD28 antibodies (0, 1, 2, 4 μg/ml) for 24 and 48 h. IL-2 and IFN-γ concentrations in supernatants were measured by ELISA. Unpaired two-tailed Student’s t-test was used to compare the results of two groups. Each sample was performed in triplicate. *P < 0.05, **P < 0.01. Unless provided in the caption above, the following copyright applies to the content of this slide: © The Author(s) Published by Oxford University Press on behalf of the British Society for Rheumatology. All rights reserved. For permissions, please article is published and distributed under the terms of the Oxford University Press, Standard Journals Publication Model ( Rheumatology (Oxford), Volume 58, Issue 4, 09 January 2019, Pages 719–728, The content of this slide may be subject to copyright: please see the slide notes for details.


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