Using RNAscope HiPlex smFISH assay, we first validated the DEGs that have been identified in human AD by other groups using snRNA-Seq or scRNA-Seq [13, 16, 24, 60, 61] [62–64], including SNCG, STMN2, CSRP1, PLP1, GLUL, CD9, C1QB, SPP1, CD63, CRYAB, and YWHAH. Those genes have been found to play important roles in the pathogenesis of AD [55, 63, 68–73]. Here, SPP1 is linked to Alzheimer disease.