Fig 1: Anti-GA or anti-PA immunoreactivity and fluorophore labelling for dipeptide repeats.(A) Dot-blotted membrane of purified poly-GA fibris, poly-GA oligomers, or poly-PA oligomers in serial dilution after staining with the respective anti-repeat antibodies (anti-GA = Proteintech 24492-1-AP; anti-PA = Proteintech 24493-1-AP). PBS indicates a control condition with no purified proteins. Graphs show quantification of Mean grey values for each corresponding condition. Data information, sample numbers: no 1 = 1 µM; no 2 = 0.5 µM; no 3 = 0.25 µM; no 4 = 0.13 µM; no 5 = 0.06 µM; no 6 = PBS only. (B) Poly-GA oligomers and fibrils labelled by ATTO-647N (molar ratio Atto:GA = 2:1) exhibit fluorescent signal and Coomassie staining after protein gel electrophoresis and fluorescent signal after dot-blot. Whereas both oligomers and fibrils exhibit fluorescence signal, the fibrils’ signal is three times higher (left). On the Coomassie gel, the quantity of monomers and fibrils is comparable (centre). (C) 5 µl at 100 µM (4.5 µg) of fibrillar poly-GA labelled by ATTO-550 (molar ratio Atto:GA = 5:1) were resolved on an acrylamide gel, and the fluorescence of Atto-550 (black signal) was recorded (left). Two samples were also analysed by dot blot on nitrocellulose membrane, and the fluorescence of Atto-550 is displayed (black signal) (right).