tissues were surgically removed and minced into small pieces (approximately 1mm3) on ice and digested by 200 U/mL Papine (Diamond) and Cysteine-HCL (Sigma). Then samples were sieved through 70 µm cell strainers, centrifuged at 300 g for 10 min, and further cleaned for red blood cells using red blood cell lysis buffer (Miltenyi Biotec). The scRNA-seq libraries was generated using the Chromium Single Cell 3′ Library Kit v3 according to the manufacturer's instructions. Single-cell suspensions from each group (e.g., CAV_brain) were loaded onto the Chromium single-cell controller to generate cell-bead droplets in the emulsion. Barcoded cDNA was amplified and fragmented for library construction according to the manufacturer's instructions. The concentrations of the final libraries were determined by Qubit (Thermo Fisher Scientific), and the fragment lengths were determined by Agilent 2100. The libraries were then pooled and sequenced on an Illumina NovaSeq 6000 platform according to the manufacturer's instructions.