Fig. 2.

Co-transplantation of miR-615 agomir with gel stimulated axonal regeneration and reduced neuronal death in affected spinal segments after avulsion/reimplantation. a, b Representative micrographs and quantitative analysis of the survival neurons around the avulsed epicenter by Nissl’s staining at low and high magnification, respectively, indicating a significant increasement of neurons in miR-615 agomir+gel rats. c Immunofluorescence photos of neurons around the avulsed site in the ipsilateral C6 segment. d Relative number of neurons were measured and counted. e, f Western blot assay and relative quantification of LINGO-1 protein demonstrated dramatic downregulation of LINGO-1 in miR-615 agomir and miR-615 agomir+gel group. The results illustrated that miR-615 negatively regulated LINGO-1. GAPDH was used as an internal control. g, h Western blot assay and relative quantification of NeuN, a neuron-specific protein, demonstrated dramatic upregulation of NeuN in miR-615 agomir and miR-615 agomir+gel group. GAPDH was used as an internal control. Data are presented as the mean ± SD (one-way analysis of variance followed by the least significant difference post hoc test). *p < 0.05, **p < 0.01, ***p < 0.001, vs. PBS group. Scale bar (a—upper row) = 500 μm, scale bar (b—lower row) = 250 μm, Scale bar (c) = 200 μm