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In a first straight forward approach a neuron-specific nuclear protein (NeuN) immunohistochemical staining of coronal brain sections was performed.
Double labeling of brain sections was performed using the rabbit anti-GPR17 polyclonal antibody (1∶100) generated as previously described [11] in combination with the selected primary antibody, in PBS 0.01 M, 0.1% Triton X-100.
Immunohistochemistry on adult or fetal human brain sections was performed using standard protocols.
The procedure for preparing brain sections was performed as described previously [ 29].
The double fluorescence in situ hybridization in fresh brain sections was performed following published protocols (Jones et al., 2007).
Immunohistochemistry on brain sections was performed following the avidin biotin peroxidase method (Vectastain ABC kit, Vector Labs, Burlingame, CA, USA).
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Histology and immunohistochemistry of brain sections were performed as described previously [16], [17].
Microdissections of rat brain sections were performed under 10x objective and target cells were collected from each rat brain on an LCM Cap (CapSure Macro Caps, Arcturus; Mountain View, CA).
All the measurements on the brain sections were performed at the approximate level of bregma −2.18.
Histomorphometric analysis of DR neurons in stained brain sections were performed in experimental and control rats; neurons in inferior colliculus (IC) served as anatomical control.
Quantitative analyses for the immunostained brain sections were performed by defining regions of interest (ROIs) within the ischemic striatum as described earlier.
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