Figure 1. BM-MSCs from SLE patients showed senescence characteristics (A,B) SA-β-gal was used to examine BM-MSC senescence. The number of SA-β-gal-positive.

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Figure 1. BM-MSCs from SLE patients showed senescence characteristics (A,B) SA-β-gal was used to examine BM-MSC senescence. The number of SA-β-gal-positive cells was obviously increased among the SLE BM-MSCs compared with that from a normal person. (C) BM-MSCs were stained with FITC-Phalloidin. Immunofluorescence showed that the F-actin distribution was abnormal in the BM-MSCs from SLE patients. (D,E) Flow cytometry results showed that the ratio of cells in G0 phase was increased in the SLE BM-MSCs compared with the normal group. *P < 0.05 compared with the normal group. NOR, normal group. From: JAK-STAT signaling mediates the senescence of bone marrow-mesenchymal stem cells from systemic lupus erythematosus patients Acta Biochim Biophys Sin (Shanghai). 2017;49(3):208-215. doi:10.1093/abbs/gmw134 Acta Biochim Biophys Sin (Shanghai) | © The Author 2017. Published by Oxford University Press on behalf of the Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com

Figure 2. JAK-STAT signaling was over-activated in BM-MSCs from SLE patients (A,B) p-JAK2 and p-STAT3 expression were higher in BM-MSCs from SLE patients compared with that from the normal group, as determined by western blot analysis. GAPDH was used as an internal control. *P < 0.05 compared with the normal group, <sup>#</sup>P < 0.05 compared with the normal group. From: JAK-STAT signaling mediates the senescence of bone marrow-mesenchymal stem cells from systemic lupus erythematosus patients Acta Biochim Biophys Sin (Shanghai). 2017;49(3):208-215. doi:10.1093/abbs/gmw134 Acta Biochim Biophys Sin (Shanghai) | © The Author 2017. Published by Oxford University Press on behalf of the Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com

Figure 3. IFN-γ increased the expression of JAK-STAT signaling and promoted cellular senescence in normal BM-MSCs (A,B) Cells were cultured in different concentrations of IFN-γ for 48 h. By determining the expression of p-STAT3, we observed that IFN-γ achieved its maximal effects at a dose of 100 ng/ml. (C) BM-MSCs were fixed and stained with SA-β-gal. The number of SA-β-gal-positive cells was increased in the IFN-γ-treated normal BM-MSCs in comparison with the normal group. (D) Immunofluorescence showed that the normal distribution of F-actin in the BM-MSCs from normal persons was disorder after stimulated with IFN-γ. (E) The flow cytometry results showed that the ratio of cells in G0 phase was increased in the IFN-γ-treated group. (F) Western blot analysis was performed to detect the protein expressions of CDK2, CyclinD1, CyclinE, and p21. *P < 0.05 compared with the normal group, <sup>#</sup>P < 0.05 compared with the normal group, <sup>%</sup>P < 0.05 compared with the normal group, <sup>&</sup>P < 0.05 compared with the normal group. From: JAK-STAT signaling mediates the senescence of bone marrow-mesenchymal stem cells from systemic lupus erythematosus patients Acta Biochim Biophys Sin (Shanghai). 2017;49(3):208-215. doi:10.1093/abbs/gmw134 Acta Biochim Biophys Sin (Shanghai) | © The Author 2017. Published by Oxford University Press on behalf of the Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com

Figure 4. AG490 and si-STAT3 slowed cellular senescence in BM-MSCs from SLE patients (A) Cells were cultured with different concentrations of AG490 for 48 h. By determining the expression of p-STAT3, we observed that AG490 achieved its maximal inhibition effects at the dose of 500 µM. (B) Determination of the knocking-down efficiencies of three STAT3-targeting siRNA oligos. (C) BM-MSCs were fixed and stained with SA-β-gal. The number of SA-β-gal-positive cells was decreased in the AG490 and STAT3-siRNA-treated SLE BM-MSCs in comparison with the SLE group. Addition of IFN-γ did not increased the ratio of SA-β-gal-positive cells in AG490-treated SLE BM-MSCs (D) Immunofluorescence showed that the disorder distribution of F-actin in the BM-MSCs from SLE patients was reversed after AG490 treatment and by si-STAT3. (E) Flow cytometry results showed that the ratio of cells in G0 phase was decreased in the AG490-treated group and si-STAT3 group. (F) Western blot analysis was performed to examine the protein expressions of CDK2, CyclinD1, CyclinE, and p21. *P < 0.05 compared with the SLE group, <sup>#</sup>P < 0.05 compared with the SLE group, <sup>%</sup>P < 0.05 compared with the SLE group, <sup>&</sup>P < 0.05 compared with the SLE group. From: JAK-STAT signaling mediates the senescence of bone marrow-mesenchymal stem cells from systemic lupus erythematosus patients Acta Biochim Biophys Sin (Shanghai). 2017;49(3):208-215. doi:10.1093/abbs/gmw134 Acta Biochim Biophys Sin (Shanghai) | © The Author 2017. Published by Oxford University Press on behalf of the Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com