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. 2019 Dec 10;14(12):e0214351. doi: 10.1371/journal.pone.0214351

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© 2019 Tanaka et al

This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

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Fig 8 — Top panels: The spinal cord at the C5 level was sliced into 5-μm-thick sections at 10 weeks after surgery and the gray matter was stained for EPO-R (original magnification ×40, scale bar = 20 μm). The expression of EPO-R in the anterior horn cells was clear in the vehicle group but weak in the EPO group. Second panels: Caspase-3 staining of spinal cord is shown. The arrows indicate caspase-3-positive nuclei, which were detected in the vehicle group but not in the EPO group. Third panels: APP staining of the gray matter of spinal cord is shown. Staining of anterior horn cells with APP was similar in both groups. Fourth panels: APP staining in the white matter of spinal cord is shown. Arrows show APP-positive cells. More APP-positive cells were observed in the vehicle group. Fifth panels: Synaptophysin staining of the white matter of spinal cord is shown. Arrows indicate synaptophysin-positive axons. In the EPO group, synaptophysin-positive axons extending from the gray matter to the white matter were clearly observed. Sixth panels: 5-HT staining of the gray matter of spinal cord is shown. Nerve fibers were clearly stained with 5-HT (indicated by arrows) in the gray matter from EPO group but not in that from vehicle group. Bottom panels: GAP-43 staining of gray matters of spinal cord is shown. The expression of positive cells was not clear in both the vehicle and EPO groups. B: The number of EPO-R-positive anterior horn cells was significantly higher in the vehicle group (P < 0.005, unpaired t-test). C: Caspase-3-positive cells were detected only in the vehicle group (P < 0.001, unpaired t-test). D: No significant difference was observed in the number of APP-positive cells in the gray matter between the two groups (P = 0.13, unpaired t-test). E: Significantly more APP-positive cells were detected in the white matter of the vehicle group (P < 0.05, unpaired t-test). F: The number of synaptophysin-positive axons was significantly higher in the high-dose EPO group (P < 0.005, unpaired t-test).