Fig. 2

Representative images of various intracellular responses to the absence of MT3. a Neuronal outgrowth induced in MT3 knockout (KO) mice. Cortical astrocytes from MT3 wild type (WT, ^+/+) and KO (^−/−) mice were used as the feeder cells. Cortical neuronal cells from ICR mice were cultured onto the astrocytes from the indicated sources. b-d Confocal images of altered localization and expression of intracellular components, such as endosomes b, Golgi c, F-actin d, in cortical astrocytes from MT3 WT and KO mice. e Fluorescence images of lysosensor-stained MT3 WT and KO cortical astrocytes. f Confocal images of altered localization and expression of vATPase Voa1 in cortical astrocytes from MT3 WT and KO mice. g-h Live-cell confocal microscopic images of WT and MT3-null astrocytes. Cortical astrocytes were exposed to 100 μm of H₂O₂ and the intracellular labile zinc was traced using a FluoZin-3 g, free zinc staining fluorescence dye, and Lysotracker h, a fluorescence dye staining of lysosomes. i-m Confocal images of altered localization and expression of intracellular proteins; Cortical astrocytes from WT and MT3-null astrocytes positive for GFP-LC3 i, fillipin and Lamp2 j, lipofuscin k, presenilin l, and GFP-mHttQ74 m. Cells were stained with filipin (green) j and lipofuscin (cyan) k against cellular lipids. Lamp2 was used to determine the cellular localization of intracellular fillipin-positive signals. GFP-mHttQ74 (green dot) was overexpressed in cortical astrocytes and its expression was compared between WT and KO of MT3. n Live-cell confocal microscopic images of WT and MT3-null astrocytes. The plasma membrane of astrocytes was stained with phalloidin, a membrane stain, and FITC-Aβ_1–42 was loaded and the intracellular uptake into astrocytes was traced. FITC-Aβ_1–42 was uptaken into cells 15 min after loading