Journal: Molecular Medicine Reports
Article Title: Lentiviral-mediated growth-associated protein-43 modification of bone marrow mesenchymal stem cells improves traumatic optic neuropathy in rats
Figure Lengend Snippet: Overexpression of GAP-43 promotes the differentiation of BMSCs into neuron-like cells. (A) Western blot analysis demonstrated that the protein expression of the GAP-43 was significantly increased in the LV5-GAP-43 group, compared with the NC and Blank groups. (B) BMSCs were observed using microscopy and it was revealed that BMSCs exhibited a neuronal phenotype following transduction with LV5-GAP-43. (C) BMSCs expressed positive specific neural markers for NSE, NF, nestin and βIII-tubulin in the LV5-GAP-43 group, determined using semi-quantitative polymerase chain reaction detection. Scale bar=50 µ m. GAP-43, growth-associated protein-43; BMSCs, bone marrow mesenchymal stem cells; NSE, neuron-specific enolase; NF, neurofilament; NC, transduced with negative control scramble lentiviral vector; LV5-GAP-43, transduced with LV5-GAP-43; M, marker.
Article Snippet: The samples were then incubated with rabbit anti-rat nestin, NeuN and GAP-43 polyclonal antibodies (Wuhan Boster Biological Technology, Ltd.) at a dilution of 1:100 in PBS at 4°C overnight, washed twice with PBS and incubated with FITC- or Cy3-labeled rabbit anti-goat IgG (H+L; Wuhan Boster Biological Technology, Ltd.) at a dilution of 1:200 in PBS at room temperature for 1 h. Finally, images were captured using an inverted fluorescence microscope (Olympus Corp.).
Techniques: Over Expression, Western Blot, Expressing, Microscopy, Transduction, Real-time Polymerase Chain Reaction, Negative Control, Plasmid Preparation, Marker