For each sample, image stacks (26 images of 0.2-μm Z-step size) were acquired for each wavelength.
Confocal image stacks (Z-stacks) with a 0.63 × 0.63 × 0.65 μm voxel size were acquired for cell volume calculations using Amira 5.4.1 (Visualization Sciences Group; Mérignac, France).
To quantify Wg fluorescence intensity in wing imaginal discs after staining, 30 Z-stack images (step size 0.5 μM) were acquired for each sample.
High-resolution T1 anatomical images (size, 256 × 256; 180 slices; size: 1 mm isotropic) were acquired for each participant before the two functional sessions.
Forty slices positioned parallel to the AC PC line and covering the whole brain (matrix size, 96 × 96; voxel size, 2.5 × 2.5 × 3.5 mm) were acquired for each volume (TR = 2.5 s; TE = 35 ms).
Three-dimensional high-resolution T1-weighted sagittal images (GE sequence IR-FSPGR, TR = 6988 ms, TI = 1100 ms, TE = 3.9 ms, flip angle = 10, voxel size = 1 × 1 × 1.2 mm3) were acquired for registration and normalization of the functional images as well as for atrophy measures and VBM analysis.
Static images were acquired for 300 s, using the same matrix, pixel size and energy settings as for the whole body images.
Whole brain, high resolution, anatomical images were acquired for all participants using a T1-weighted sequence with a voxel size of 1 × 1 × 1 mm.
For each measurement set, scans were acquired for duration of around 1 min. The number of scans acquired depends on the scan rate which in turn depends on the waveform scan resolution, the window size, and the Integration size.
Radioisotopic data were acquired for 180 minutes.
