Fig. 2

The UTX^−/− ECs promote macrophages polarization toward M2 subtype after SCI. A Immunofluorescence analysis of the UTX (red) expression in CD31⁺ endothelial cells (green) in sham, UTX^f/f, and UTX^−/− mice at 7 days after SCI. Scale bar, 20 μm. B Statistical analysis of the mean fluorescence intensity of UTX in A. C Immunofluorescent co-staining of the M2 marker (CD206, red) and F4/80⁺ macrophages (green) in sham, UTX^f/f and UTX^−/− mice at 7 days post-SCI. Scale bars, 20 μm. D Statistical analysis of mean fluorescence intensity of CD206 in C. E Immunofluorescence analysis of the M1marker (iNOS, red) expression in F4/80⁺ macrophages (green) in sham, UTX^f/f, and UTX^−/− mice at 7 days after SCI. Scale bars, 20 μm. F Statistical analysis of the mean fluorescence intensity of iNOS in E. G–J Relative mRNA expression of M1 macrophage markers (iNOS2, TNF-ɑ, CD86 and CD38). K–N Relative mRNA expression of M2 macrophage markers (Arg-1, IL-10, CD206 and Ym-1). O Representative scatter plots of macrophage phenotypes in sham, UTX^f/f, and UTX^−/− mice at day 7 after SCI. Cells were immunolabeled with antibody-fluorophore coupled antibodies to CD45-APC-Cy7, CD11b-PerCP-Cy5.5, F4/80-FITC, CD206-APC, and CD11c-PE. Macrophages were defined as CD45^highCD11b⁺, F4/80⁺ cells were defined as mature macrophages, and CD206 and CD11c defined macrophage phenotypes, M0 macrophages (CD11c⁻CD206⁻), M1 macrophages (CD11c⁺CD206⁻) and M2 macrophages (CD11c⁻CD206⁺). The numbers in the gates refer to the percentage of positive cells for each marker. P The percentage of macrophages (CD45^highCD11b⁺) in O. Q The percentage of F4/80⁺ mature macrophage cells in the CD45^highCD11b⁺ gate (macrophages). R, S The percentage of M1 macrophages (CD11c⁺CD206⁻) and M2 macrophages (CD11c⁻CD206⁺) cells in the CD45^highCD11b⁺F4/80⁺ gate (mature macrophages). ^nsP > 0.05, *P < 0.05, **P < 0.01, compared with corresponding control mice. n = 5/per group