Fig. 8.

Validation of identified representative DEGs associated with AD pathology at the single-cell level using RNAscope smFISH. (A) Representative RNAscope images (left panel) and their corresponding post-IF staining of Aβ, AT8, and GFAP (right panels) in layer V for cells distal to and proximal to pathological spots. All images from the same cell were aligned and registered by imageJ (see details in Methods). (B) RNAscope probes against human SLC1A3, KIF5A, SNCG, STMN2, CSRP1, PLP1, GLUL, PAQR6, CD9, C1QB, SPP1, CD63, CRYAB, and YWHAH in microglia (P2RY12⁺/C1QB⁺), astrocytes (GFAP⁺), neurons (RBFOX3⁺), and oligodendrocytes (MBP⁺) were quantified and compared within three AD cases. For the quantification and definition of proximal to pathology/distal to pathology, please find the details in Methods. For quantification, we identified 20–50 microglia, astrocytes, and oligodendrocytes, and 60–90 neurons from each case. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001 (Mann–Whitney test, in proximal to pathology vs distal to pathology). Scale bar, 50 µm. See high-resolution RNAscope images at: https://bmbls.bmi.osumc.edu/scread/stofad-2