Previous studies of neuron survival by the Priestley lab have confirmed with toluidine blue histochemistry that NeuN immuno-reactivity is a reliable marker of neuron survival after spinal cord injury [34].
In addition, we assume colocalization with the NeuN, known as the marker of neuron in TUNEL-positive cells.
The levels of neuron-specific enolase, a marker of neuron death, are also reduced in patients treated with hypothermia following CPR [ 26].
Compared to nonimmunized SJL, EAE-SJL mice demonstrated more staining for β1i and less extensive staining for NeuN protein (Fig. 2 A, B ), a marker of neurons, linking neuronal damage to IP activity (19).
NAA/Cr is generally considered to be a marker of neurons, being reduced in conditions where there is neuronal loss or dysfunction [19, 20].
Identification of neurons was performed with a marker of neurons, microtubule associated protein 2 (1∶1000, Chemicon, Temecula, CA, USA).
To determine the cell type with reporter activation, we double stained for elav, a marker of neurons, or repo, a glial marker.
We stained cells for Tuj1, a marker of neurons, and determined the proportion of Tuj1-positive cells.
Immunofluorescence staining showed the presence of Tau protein, the specific marker of neurons (Fig. 4b, 2), and GFAP, the specific marker of glia cells (Fig. 4b, 3) in the cells after neurogenic induction.
Deparaffinized sections were incubated with 1% bovine serum and then reacted with primary antibodies against Nmnat3 (1 : 100; ProSci Incorporated, Poway, CA, USA), neurofilament-L (a marker of neurons; 1 : 100; DAKO Corporation, Carpinteria, CA, USA), or Thy-1 (a marker of RGCs; 1 : 100; Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA) diluted in 1% bovine serum overnight at 4 °C.
As controls in the biotinylation assay we analyzed neural cell adhesion molecule (NCAM), a cell surface marker of neurons, and actin, a cytoskeletal protein located inside cells, As expected, the majority of NCAM molecules were biotinylated, whereas actin was mainly unbiotinylated, indicating that cell surface proteins were efficiently biotinylated.
