Mice were euthanized and exsanguinated by transcardial perfusion with PBS, after which brains were rapidly removed and rinsed in ice-cold PBS. Tissues were cryoprotected in 30% sucrose, embedded in OCT, and stored at −80 °C. Coronal brain cryosections (10 μm thickness) were cut and mounted onto Visium Spatial Gene Expression Slides (10x Genomics) containing spatially barcoded capture spots. Sections were subjected to hematoxylin and eosin (H&E) staining and imaging prior to downstream transcriptomic processing. ollowing imaging, tissue sections were permeabilized to release mRNA, which hybridized to spatially barcoded oligonucleotides on the Visium capture areas. Reverse transcription was performed in situ to generate spatially barcoded cDNA, which was subsequently collected, amplified, and processed into sequencing libraries according to the manufacturer's protocol. Libraries were sequenced on an Illumina NovaSeq 6000 platform (S4 flow cell, XP chemistry), enabling whole-transcriptome profiling with spatial resolution at ~55 μm spot size.