Neurones, astrocytes, and capillary endothelial cells were isolated from separate sections of the same frozen blocks of lateral temporal cortex, using rapid immuno-laser capture microdissection (LCM), which preserves RNA quality sufficiently for array analysis, as described previously [53, 63]. This method produces cell-type enriched, but not pure, samples. Toluidine blue-stained neurones, glial fibrillary acidic protein (GFAP)-stained astrocytes, and collagen IV-stained endothelial cells were isolated from each case using the PixCell II LCM System (Arcturus Engineering, Mountain View, CA, USA) (Table 2). Total RNA was extracted using PicoPure® RNA Isolation Kits (Arcturus BioScience, Mountain View, CA, USA), according to the manufacturer’s protocol, with typical yields of 86.0 ng (SD 2.5 ng; range 52.3–133.7 ng), 148.4 ng (SD 2.8 ng; range 101.0–186.8 ng), and 141.2 ng (SD 8.0 ng; range 31.6–311.4 ng) of total RNA from neurones, astrocytes, and endothelial cells, respectively. The concentration and purity (A260/A280 ratio) of the total RNA were measured using the NanoDrop 1000 Spectrophotometer (Thermo Scientific, Wilmington, DE, USA). The quality of the total RNA was assessed using the 2100 Bioanalyser with RNA 6000 Pico LabChip Kits (Agilent, Palo Alto, CA, USA).

Antibody source and specificity

AGE, advanced glycation end-product; COL4, collagen IV; COX5b, cytochrome C oxidase subunit 5B; EDTA, ethylenediaminetetraacetic acid; FOXO3a, forkhead box O3; GFAP, glial fibrillary acid protein; IGF-1Rβ, insulin-like growth factor 1 receptor beta; IgG, immunoglobulin G; MW, microwave; N/A, not applicable (frozen tissue); NDUFb6, NADH:ubiquinone oxidoreductase subunit B6; O/N, overnight; p53, tumour protein p53; PC, pressure cooker; RT, room temperature; TB, toluidine blue; TGFβ1, transforming growth factor beta 1; TSC, trisodium citrate; γH2AX, gamma H2A histone family member X

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