Mouse anti-NeuN conjugated to Alexa Fluor 488 (1 100), mouse anti-GFAP conjugated to Alexa Fluor 488 (1 100) or rat anti-CD68 (1 100) were applied as marker for neuron, astrocyte or microglial cells, respectively.
Therefore, we used NeuN (RBFOX3) as marker for neuron density and found a significant reduction in NeuN in the AD patients compared with NDC above 72 years of age (p = 0.006, Figure 2B), whereas no differences between these groups in kif21b expression were found.
However, a very small numbers of the cells expressed GFAP, a marker for astrocytes, O4, a marker for immature oligodendrocytes, and βIII-tubulin, a marker for neurons.
Very small fractions, typically 0.5% or less, of the cells expressed GFAP (Figure 2 B), a marker for astrocytes, βIII tubulin, a marker for neurons, and O4, a marker for immature oligodendrocytes.
These neurons express both the sex-specific transcripts of the fruitless gene (fru+), a marker for neurons likely to have sex-specific functions, and pickpocket (ppk +), which is believed to encode a marker for proprioceptive neurons [ 14 17].
A second series was stained for NeuN, which is a selective marker for neurons that helps visualize the extent of any lesion.
Most of these were still highly positive for nestin (a marker for neural stem/progenitor cells), while a few cells appeared that were doubly positive for nestin and either GFAP (marker for astrocytes) or β-tubulin III (marker for neurons).
A morphometric analysis of SCN in aged female marmosets compared with adult animals showed a significant decrease in the number of neurons stained with Nissl and labeled immunohistochemically for NeuN, a specific marker for neurons.
As the RNAi vector had a green fluorescent protein (GFP) expression cassette within it, the cells with green fluorescence were therefore transfected cells; microtubule-associated protein 2 (MAP2; red) was used as a marker for neurons.
All of 8 coronal sections prepared were reacted with a mouse monoclonal antibody against NeuN (1 500) (CHEMICON, Billerica, MA, USA), a marker for neurons, followed by an Alexa Fluor 488-conjugated secondary antibody (1 500) (Invitogen, Carlsbad, CA, USA).
Furthermore, we did not observe any significant changes in the pattern of Tuj1 staining, a marker for neurons, nor any increase in expression of Tbr2, a marker for differentiating intermediate progenitor cells (Kowalczyk et al, 2009), 24 h after electroporation (Supplementary Figure S1D and E).
