Article Figures & Data
Figures
- Figure 1.
Tight junction protein levels in the spinal cords of G93A SOD1 mice via Western blotting. Media-treated ALS mice showed significant reductions of ZO-1 (A, B), occludin (C, D), and claudin-5 (E, F) protein levels versus controls. Although elevated levels of analyzed tight junction proteins were noted in ALS mice receiving hBM34+ cells, significant differences between media and these cell-treated mice were not found. In contrast, significant increases of ZO-1 (A, B), occludin (C, D), and claudin-5 (E, F) proteins were determined in ALS mice after hBM-EPC transplantation versus media-treated animals. For each evaluated protein (B, D, F), the band density was normalized to GAPDH signal and fold signal changes relative to levels of controls were presented; *p < 0.05, **p < 0.01.
- Figure 2.
Immunohistochemical staining for pericytes (CD13) and laminin in cervical spinal cord capillaries of G93A SOD1 mice. A, Double immunohistochemical staining in control mice showed high CD13 (a, red, arrowhead) and laminin (a’, green, arrowhead) immunoexpressions in cervical spinal cord capillaries. In media-treated mice, substantially decreased immunostaining of CD13 (b, red, arrowhead) and laminin (b’, green, arrowhead) in capillaries was noted. Mice treated with hBM34+ cells demonstrated higher CD13 (c, red, arrowhead) and laminin (c’, green, arrowhead) immunoexpressions in cervical spinal cord capillaries versus media-injected mice. Similarly, hBM-EPC-treated mice displayed increased immunoexpressions of CD13 (d, red, arrowhead) and laminin (d’, green, arrowhead) within capillaries of cervical spinal cords. Cervical spinal cord images (a’’–d’’) are merged with DAPI. Scale bar in all images is 20 μm. B, Quantitative analysis of CD13 and laminin immunoexpressions in cervical spinal cord capillaries showed significant decreases of both antigens in media-injected mice versus controls. In hBM34+ and hBM-EPC-treated mice, significant increases of CD13 and laminin immunoexpressions were determined compared with media-treated animals. Also, hBM-EPC-treated mice displayed significantly higher antigen levels versus mice treated with hBM34+ cells; **p < 0.01.
- Figure 3.
Immunohistochemical staining for pericytes (CD13) and laminin in lumbar spinal cord capillaries of G93A SOD1 mice. A, Similar to the cervical spinal cord, double immunohistochemical staining in control mice showed high CD13 (a, red, arrowhead) and laminin (a’, green, arrowhead) immunoexpressions in lumbar spinal cord capillaries. A substantial decrease of immunostaining of CD13 (b, red, arrowhead) and laminin (b’, green, arrowhead) in capillaries was noted in media-treated ALS mice. Mice receiving hBM34+ cells showed higher CD13 (c, red, arrowhead) and laminin (c’, green, arrowhead) immunoexpressions in lumbar spinal cord capillaries versus media-treated mice. hBM-EPC-treated mice also displayed increased immunoexpressions of CD13 (d, red, arrowhead) and laminin (d’, green, arrowhead) within capillaries. Lumbar spinal cord images (a’’–d’’) are merged with DAPI. Scale bar in all images is 20 μm. B, Quantitative analysis of CD13 and laminin immunoexpressions in lumbar spinal cord capillaries showed significant decreases of both antigens in media-treated mice versus controls. Immunoexpressions of CD13 and laminin significantly increased in hBM34+ and hBM-EPC treated versus media-treated mice. However, hBM-EPC-treated mice demonstrated significantly higher antigen levels versus mice treated with hBM34+ cells; **p < 0.01.
- Figure 4.
Fluorescent staining for F-actin and CD31 (PECAM-1) in cervical spinal cord capillaries of G93A SOD1 mice. A, Double fluorescent staining in control mice showed continuous F-actin (a, green, arrowhead) and CD31 (a’, red, arrowhead) expressions in cervical spinal cord capillaries. In media-treated mice, substantially decreased fluorescent staining of F-actin (b, green, arrowhead) and CD31 (b’, red, arrowhead) was determined. Mice treated with hBM34+ cells demonstrated continuous F-actin (c, green, arrowhead) and CD31 (c’, red, arrowhead) expressions almost identical to controls in cervical spinal cord capillaries. Similarly, hBM-EPC-treated mice displayed near circumferential fluorescent expressions of F-actin (d, green, arrowhead) and CD31 (d’, red, arrowhead) within capillaries. Images (a’’–d’’) are merged with DAPI. Scale bar in all images is 20 μm. B, Quantitative analysis of F-actin and CD31 fluorescent expressions in cervical spinal cord capillaries showed significant decreases of both antigens in the media-treated mice versus controls. In hBM34+ and hBM-EPC-treated mice, significant increases of F-actin and CD31 expressions were determined compared with media-treated animals. Also, hBM-EPC-treated mice displayed significantly higher antigen levels versus mice treated with hBM34+ cells; **p < 0.01.
- Figure 5.
Fluorescent staining for F-actin and CD31 (PECAM-1) in lumbar spinal cord capillaries of G93A SOD1 mice. A, Similar to cervical spinal cord findings, double fluorescent staining in control mice showed continuous F-actin (a, green, arrowhead) and CD31 (a’, red, arrowhead) expressions in lumbar spinal cord capillaries. A substantial decrease in fluorescent staining of F-actin (b, green, arrowhead) and CD31 (b’, red, arrowhead) was noted in media-treated mice. ALS mice treated with hBM34+ cells demonstrated continuous F-actin (c, green, arrowhead) and CD31 (c’, red, arrowhead) fluorescent expressions in lumbar spinal cord capillaries. hBM-EPC-treated mice also showed increased expressions of F-actin (d, green, arrowhead) and CD31 (d’, red, arrowhead). Images (a’’–d’’) are merged with DAPI. Scale bar in all images is 20 μm. B, Quantitative analysis of F-actin and CD31 fluorescent expressions in lumbar spinal cord capillaries demonstrated significant decreases of both antigens in media-treated mice versus controls. Fluorescent expressions of F-actin and CD31 significantly increased in hBM34+ and hBM-EPC treated versus media-injected mice. Of note, hBM-EPC-treated mice displayed significantly higher antigen levels versus ALS mice treated with hBM34+ cells; **p < 0.01.
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