Fig. 3

Induced neuronal differentiation and neuroprotection by multiple injections of hMSC at the lesional site. a Schematic of multiple intravenous injections of hMSC in contusive acute spinal cord injury (SCI). Animals were injected with hMSC (1’ MSC = single injection at 24 h post injury, 2’MSC = Dual injection at 24 h and one week post injury). b IF images show merged 4′,6-diamidino-2-phenylindole and hMSC at the lesional site at six weeks. c Multifluorescent, confocal images showing the expression of NeuN and hMSC in the gray matter. d Multi-fluorescent confocal images showing the expression of NGF and hMSC around the gray matter. e MAP-2 positivity indicates the lesional site and gray matter around and within hMSCs in trauma-induced gliosis at the posterior lesional site at six weeks. f WB images of NeuN, NGF and MAP-2 expression in lesional site segments (\(\sim\) 1 cm). g WB results of NeuN expression in the lesional site segments. h WB results of NGF expression in the lesional site segments. i WB results of MAP-2 expression in the lesional site segments. Full-length western blot images are presented in Additional file 7: Fig. S7. j BBB scales in the 1’MSC and 2’MSC groups are significantly higher than that in the PBS group. Data are presented as mean ± SEM. Statistical significance was estimated using Kruskal–Wallis test with post hoc analysis and Mann–Whitney (†) test with least significant difference post hoc analysis (*); *, † P < 0.05. (WB analysis: PBS n = 5, 1’MSC n = 4 and 2’MSC n = 5, BBB locomotor scales: PBS n = 7, 1’MSC n = 5, 2’MSC n = 6) Scale bars = 20 μm. hMSC, human mesenchymal stem cells; IF, immunofluorescence; BBB, Basso–Beattie–Bresnahan; WB, Western blotting