Journal: Acta Neuropathologica Communications

Article Title: Tunneling nanotube (TNT)-mediated neuron-to neuron transfer of pathological Tau protein assemblies

doi: 10.1186/s40478-016-0386-4

Figure Lengend Snippet: Tau is associated with TNTs in neuronal CAD cells. Cells were plated in Lab-Tek chamber slides and co-infected with LVs encoding mCherry-Actin ( red ) and V5-hTau1N4R ( green ). Neuronal CAD cells were processed for immunocytochemistry analysis using anti-V5 antibodies visualized with an Alexa 488-labeled secondary antibody ( green ) and anti-acetylated-tubulin visualized with an Alexa 647-labeled secondary antibody ( white ). Nuclei were labeled with DAPI ( blue ). Cells were imaged with an inverted laser-scanning confocal microscope using a 40× oil-immersion lens (NA 1.3), and the images were processed using ZEN and ImageJ. A focal plane was collected for each specimen. Scale bars: 10 μm. TNTs ( white arrows ), which are not always bound to the dish, are shown in enlargements

Article Snippet: Uptake and transfer of hTau1N4R fibrils- Neuronal CAD cells were plated overnight at 60,000 cells per well in poly-D-lysine-coated Lab-Tek four-well chamber slides (Becton Dickinson).

Techniques: Infection, Immunocytochemistry, Labeling, Microscopy

Journal: Acta Neuropathologica Communications

Article Title: Tunneling nanotube (TNT)-mediated neuron-to neuron transfer of pathological Tau protein assemblies

doi: 10.1186/s40478-016-0386-4

Figure Lengend Snippet: Exogenous Tau fibrils are found within TNTs. CAD neuronal cells ( a ) or primary neurons ( b ) were plated in Lab-Tek chamber slides and infected with LVs encoding mCherry-Actin (red). At 48 h post-infection, cells were incubated for six hours with extracellular ATTO 488-hTau1N4R fibrils/liposomes preparation ( green ). Cells were processed for immunostaining analysis using an anti-acetylated tubulin visualized with an Alexa 647-labeled secondary antibody ( white ). Nuclei were labeled with DAPI ( blue ). TNTs containing Tau are shown in enlargements (white arrows). For acquisition, a focal plane was collected for specimen. Images were acquired with an inverted laser-scanning confocal microscope using a 40× oil-immersion lens (NA 1.3) and processed using the programs ZEN and ImageJ. Scale bars: 10 μm

Article Snippet: Uptake and transfer of hTau1N4R fibrils- Neuronal CAD cells were plated overnight at 60,000 cells per well in poly-D-lysine-coated Lab-Tek four-well chamber slides (Becton Dickinson).

Techniques: Infection, Incubation, Immunostaining, Labeling, Microscopy