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Confocal microscopic observations of fly brains were performed using a Zeiss LSM 510 laser scanning confocal microscope (Carl-Zeiss, Germany).
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High-resolution (66 × 66 × 100 µm; scan time = 96 min) ex vivo MRI of brains was performed using a 3-Tesla (T) MR scanner interfaced with customized gradient and radiofrequency coils.
PET/CT of the brain was performed using a GE Discovery VCT PET/CT Scanner General Electric Medical Systemss, Pewaukee, WI, USA).
3D reconstruction of each brain was performed using customized procedures ([57] and File S1).
Then, tissue segmentation from the transformed images to the gray matter, white matter, CSF space, and non-brain was performed using the SPM2 default segmentation procedure.
Gentle mechanical desheathing of the exposed brain was performed using fine forceps.
Sex specific DNA methylation analysis in brain was performed using published dataset (GEO Accession No: GSE15745) [ 29].
Low molecular weight (LMW) RNA enrichment from whole mouse brain was performed using mirVana™ miRNA Isolation Kit (Ambion), according to the manufacturer's protocol.
Immunohistochemistry to detect isoQC and CCL2 in human brain was performed using the rabbit anti-isoQC antiserum 3285 1 5000) and the mouse monoclonal CCL2 antibody MAB2791 1 5000), respectively.
Immunohistochemistry to detect QC in human brain was performed using a commercially available mouse anti-human QC antiserum (A01; Abnova; 1 500).
Brain MRIs were performed using a Siemens Sonata equipment, Erlangen, Germany (1.5 T).
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