To solve this issue, we chose RNAscope smFISH in combination with post-IF of Aβ plaques, NFTs and neuropil threads (AT8+), and GFAP to validate DEGs associated with these AD pathologies at the single-cell level using three AD (Braak III-IV) and three CT cases (Braak I-II). The gene discussed is GFAP; the disease is Alzheimer disease.