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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).
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].
Immunohistochemistry to detect QC in human brain was performed using a commercially available mouse anti-human QC antiserum (A01; Abnova; 1 500).
Low molecular weight (LMW) RNA enrichment from whole mouse brain was performed using mirVana™ miRNA Isolation Kit (Ambion), according to the manufacturer's protocol.
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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.
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.
Confocal microscopic observations of fly brains were performed using a Zeiss LSM 510 laser scanning confocal microscope (Carl-Zeiss, Germany).
Brain extraction was performed using the brain extraction tool in FSL (Smith 2002).
The 3D anatomical images were transformed to match the dimensions of the functional scans using FSLSWAP and brain extraction was performed using the FSL Brain Extraction Tool (BET) (fractional intensity threshold: 0.6, threshold gradient of −0.1, and robust brain center estimation).
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