nearest 0.1 mg. The dissection includes a small part of the subiculum adjacent to CA1 and occasionally a small strand of the fimbria. For each sample, 100 μg of extracted proteins were digested with LysC (1:100, w/w) for 3 h. Samples were diluted four times with 50 mM HEPES followed by trypsin digestion (1:50, w/w) overnight at room temperature. After digestion, the samples were acidified with trifluoric acid to a pH<2 and desalted and dried in a speed vacuum. Peptides were labelled with six-plex TMT reagents (Thermo Scientific) as recommended by the manufacturer. Following labelling, the TMT-labelled samples (TMT126-TMT131) were mixed equally to generate a digest mixture which was further fractionated by high pH reverse phase liquid chromatography. Ten fractions are collected and further analysed by low pH reverse phase liquid chromatography-tandem mass spectrometry (MS/MS).
