Fig. 3.
Identifcation of α-synuclein aggregates by thioflavine S staining and by electron microscopy. A–C, Treatment of cells expressing A53T α-synuclein (A), wild-type α-synuclein (B), or untransfected cells (C) with 10 mm FeCl2 and 100 μm H2O2 for 72 hr.D–F, Treatment of cells expressing A53T α-synuclein (D), wild-type α-synuclein (E), or untransfected cells (F) with 10 mm FeCl2 for 72 hr. G, H, Inclusions are evident by electron microscopy. BE-M17 cells stably transfected with A53T α-synuclein were treated with 10 mm FeCl2 and 100 μm H2O2 for 72 hr and then examined by electron microscopy. The aggregates that formed under these conditions were long fibrils with a diameter of ∼10 nm (G, magnification of 35,000×). Although treatment with 10 mm FeCl2 and 100 μmH2O2 was toxic to many cells, some cells containing aggregates had both fibrillar deposits (triangle) and organelles that were intact (arrow), suggesting that aggregation can occur in living cells (H, magnification of 20,000×).Arrows point to thioflavine S-positive aggregates.