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The early frames were manually coregistered to the mouse brain template available in PMOD for each mouse PET image.
After spatial normalization to a mouse brain template
The final transformation matrix was then applied to the whole dynamic PET sequence for each mouse, resulting in PET images that were coregistered to the mouse brain template in PMOD.
PMOD v3.3 software (PMOD Technologies Ltd ,Zurich, Switzerland) was used for the coregistration of the PET images to a 3-D digital T2-weighted MRI mouse brain template incorporated in PMOD.
Figure 2a shows parametric BPND images of C-DED PET illustrated by coronal sections coregistered to a 3-D T2-weighted MRI mouse brain template for representative APPswe and wt mice.
Spatial normalization of the mouse brain parametric maps generated from the diffusion data (including FA and FD maps) and the morphological T2*-and T2-weighted images was performed using Matlab-script, which registered the brain maps from all investigated animals on a previously created mouse brain template.
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In vivo SPECT image (summed frames between 50 and 80 min of scan), co-registered with a mouse brain MRI template, obtained from one mouse bearing the GL26 tumor in the coronal (A), sagittal (B), and axial (C) planes.
For VOI analysis, processed images were subsequently co-registered with a predefined mouse brain VOI template (encompassing all relevant brain regions) aligned with the aforementioned Mirrione CT/MRI atlas and tracer uptake values were extracted for each delineated VOI.
GluA4S cDNA in pBluescript, kindly donated by Dr Andres Buonanno (NICHHD-NIH, Bethesda, MD), and mouse brain cDNA served as templates for the PCR reactions.
In order to clone mouse WNK2 (NCBI accession number NP_083637; residues 1650 1808) and mouse WNK3 (NCBI accession number: XP_914679; residues 1145 1508), reverse transcription PCR was carried out using mouse brain RNA (Stratagene) as a template and the SuperScript III 1 Step kit (Invitrogen).
An image of a normal mouse brain was selected as the reference template.
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