Fig. 7

EPC-EXOs-derived miR-222-3p promotes anti-inflammatory macrophages via the SOCS3/JAK2/STAT3 axis in vitro and in vivo and improves the outcome of spinal cord injury. A Western Blotting analysis of the expression levels of key proteins of the SOCS3/JAK2/STAT3 signaling pathway (SOCS3, JAK2, p-JAK2, STAT3, and p-STAT3) after EPC-EXOs, FLLL32, and EPC-EXOs + FLLL32 interventions in BMMs. B, C Statistical analysis of the expression levels of SOCS3, p-JAK2/JAK2 and p-STAT3/STAT3, n = 3, ^nsP > 0.05, *P < 0.05, **P < 0.01. D Western Blotting analysis of the expression levels of iNOS and Arg-1 in the BMMs treated with EPC-EXOs, EPC-EXOs Mimic, EPC-EXOs Inhibitor and EPC-EXOs FLLL32. E, F Statistical analysis of the expression levels of iNOS and Arg-1 in each group, n = 3, **P < 0.01. G Representative immunofluorescence images of the groups: Vehicle, EPC-EXOs, EPC-EXOs Mimic, EPC-EXOs Inhibitor and EPC-EXOS FLLL32 interfering with CD86/CD206 (red) and DAPI (blue) staining of F4/80⁺ macrophages (green). Scale bar: 50 μm. H, I Statistical analysis of the proportion of CD86⁺F4/80⁺ cells or CD206⁺F4/80⁺ cells to total F4/80⁺ cells, n = 3, **P < 0.01. J, K Immunofluorescence images of F4/80⁺ macrophages (green), CD86⁺/CD206⁺ (red) and DAPI (blue) staining of spinal cord injury sections of mice representative of the groups: Control, EPC-EXOs Mimic, EPC-EXOs Mimic FLLL32 and FLLL32 at 7- and 14- days post-injury (dpi) in wild-type mice. Scale bar: 20 μm. L–O Immunofluorescence results were analyzed by ImageJ, GraphPad, and SPSS. n = 3, ^nsP > 0.05, *P < 0.05, ***P < 0.001. P, S Western Blotting detected the expression levels of iNOS and Arg-1 in macrophages in the region of injury at 7 and 14 days post spinal cord injury in the groups: Control, EPC-EXOs Mimic, EPC-EXOs Mimic FLLL32 and FLLL32. Q–U Statistical analysis of the expression levels of iNOS and Arg-1, n = 3, ^nsP > 0.05, **P < 0.01, ***P < 0.001. V BMS primary score over time post-SCI in Sham, Vehicle, EPC-EXOs, EPC-EXOs Mimic and EPC-EXOs Mimic FLLL32 group, n = 5, ^nsP > 0.05, **P < 0.01. W Representative images of motor-evoked potential (MEP) in Sham, Vehicle, EPC-EXOs, EPC-EXOs Mimic and EPC-EXOs Mimic FLLL32 group at 28 days post-SCI. X, Y Quantification of W, n = 6, *P < 0.05