Transcription Factor SOX10 Improves Migration and Homing of MSCs After Myocardial Infarction by Upregulating CXCR4

Figures

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  Figure 1. Isolation and culture of human mesenchymal stem cells (MSCs). (a) Flow cytometry analysis of MSC surface markers. (b) ORO staining of MSCs (magnification, ×200).

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  Figure 2. CXCR4 promotes invasion and migration of MSCs. (a) RT-qPCR detection of CXCR4 expression in MSCs transfected with interference and overexpression lentiviruses. (b) Western blot analysis of CXCR4 expression in MSCs. (c) Wound healing assay to assess MSC migration ability. (d) Transwell assays to measure MSC invasion ability. MSC, mesenchymal stem cell. ⁣∗∗p < 0.01, ⁣∗∗∗p < 0.001, and ⁣∗∗∗∗p < 0.0001.

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  Figure 3. MSCs overexpressing CXCR4 improve cardiac function in a rat MI model. (a) Echocardiography assessing cardiac function 4 weeks after MI induction and MSC transplantation. (b) Quantification of LVEDD. (c) Quantification of LVESD. (d) Quantification of LVEF. (e) Quantification of LVFS. ⁣∗p < 0.05 compared to the NC group, #p < 0.05 compared to the Ctrl group, △p < 0.05 compared to the Sham group, □p < 0.05 compared to the MI group. MSC, mesenchymal stem cell.

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  Figure 4. CXCR4 promotes MSC homing to alleviate MI in ratsa. (a) Masson staining of myocardial fibers in each group (n = 5/group) (magnification, ×200). (b) Fluorescence microscopy (magnification, ×400) to measure MSC homing ability in rat myocardium (n = 5/group). ⁣∗p < 0.05 compared to the NC group, #p < 0.05 compared to the Ctrl group, □p < 0.05 compared to the MI group. MI, myocardial infarction; MSC, mesenchymal stem cell.

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  Figure 5. Knockdown of CXCR4 reverses SOX10-induced effects on MSC migration and invasion. (a) JASPAR database predicts binding sites between SOX10 and CXCR4. (b, c) Dual-luciferase reporter and ChIP assays were performed to detect the SOX10 binding to the CXCR4 promoter in MSCs. (d–f) The expression of CXCR4 by RT-qPCR and Western blotting. (g) Wound healing detected the cell migration ability. (h) Transwell assays detected the cell invasion ability. MSC, mesenchymal stem cell. ⁣∗∗∗p < 0.001 and ⁣∗∗∗∗p < 0.0001.

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  Figure 6. Knockdown of CXCR4 reverses the beneficial effects of SOX10-overexpressing MSCs in a rat MI model. (a) Echocardiography determining cardiac function. (b) Quantification of LVEDD. (c) Quantification of LVESD. (d) Quantification of LVEF. (e) Quantification of LVFS. LVEDD, LV end-diastolic dimensions; LVEF, LV ejection fraction; LVFS, LV fractional shortening; MI, myocardial infarction; MSC, mesenchymal stem cell. ⁣∗p < 0.05, ⁣∗∗p < 0.01, and ⁣∗∗∗p < 0.0001.

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  Figure 7. CXCR4 knockdown reverses MSC homing ability. (a) Masson staining of myocardial fibers in each group (n = 5/group) (magnification, ×200). (b) Fluorescence microscopy (magnification, ×400) to assess MSC homing in rat myocardium (n = 5/group). MSC, mesenchymal stem cell. ⁣∗∗∗p < 0.001 and ⁣∗∗∗∗p < 0.0001.