Exercise-Intervened Endothelial Progenitor Cell Exosomes Protect N2a Cells by Improving Mitochondrial Function

Figures

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  Figure 1. Exercise intervention improved the incorporation efficiency of EPC-EXs isolated from hypertensive transgenic mice by N2a cells. (A) representative images showing the incorporation of EX (labeled with PKH26, a red fluorescence) uptake by N2a cells at 8 h, 16 h, and 24 h incubation. Magnification: 200×. (B) summarized data showing the EPC-EX incorporation by N2a cells. Blue: DAPI. Red: PKH26 labeled EXs. * p < 0.05, vs. R-EPC-EX in 16 h; + p < 0.05, vs. R-EPC-EX in 24 h. Data expressed as mean ± SD, n = 3/group.

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  Figure 2. Profiling analysis of exosomal miRs and qRT-PCR of miR-27a in EPC-EXs of exercised and sedentary hypertensive mice. (A) representative miR profiling graph showing the changes of miRs in EPC-EXs. MiRs with down- or up-regulated >3-fold are listed. (B) qRT-PCR verified the miR-27a level in EPC-EXs. * p < 0.05, vs. R-EPC-EX. Data are expressed as mean ± SD, n = 3/group.

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  Figure 3. Exercise-intervened EPC-EXs raised miR-27a level, improved the cell survival ability, and reduced ROS overproduction in N2a cells challenged by Ang II plus hypoxia. (A) miR-27a level in N2a cells co-cultured with or without R-EPC-EXs. (B) summarized data showing the N2a cell viability assessed by MTT assay. (C) representative images of N2a cells of DHE staining (red fluorescence). Scale bar: 100 μm. (D) summarized data of DHE staining assay for ROS production in N2a cells. * p < 0.05, vs. con (no Ang II + hypoxia); + p < 0.05, vs. Ang II + hypoxia only; # p < 0.05, vs. R-EPC-EX; ^ p < 0.05, vs. R-EPC-EXET. Data expressed as mean ± SD, n = 3/group.

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  Figure 4. Exercise-intervened EPC-EXs regulated the MMP in N2a cells challenged by the Ang II plus hypoxia. (A) representative images show the JC-1 staining in N2a cells. Enlarged images are from the dashed box of the merged images. Scale bar: 50 μm. (B) summarized data of JC-1 stain. * p < 0.05, vs. con (no Ang II + hypoxia); + p < 0.05, vs. Ang II + hypoxia only; # p < 0.05, vs. R-EPC-EX; ^ p < 0.05, vs. R-EPC-EXET. Data expressed as mean ± SD, n = 3/group.

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  Figure 5. Exercise-intervened EPC-EXs regulated the expressions of Nox4 and cytochrome c in Ang II plus hypoxia-challenged N2a cells. (A–C) representative bands, and summarized data of cytochrome c and Nox4 levels in N2a cells. * p < 0.05, vs. con (no Ang II + hypoxia); + p < 0.05, vs. Ang II + hypoxia only; # p < 0.05, vs. R-EPC-EX; ^ p < 0.05, vs. R-EPC-EXET. Data expressed as mean ± SD, n = 3/group.