Skip to main content
Fig. 4 | Acta Neuropathologica Communications

Fig. 4

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

Fig. 4

Tau is found in TNTs in primary neurons. a Neurons were infected with LVs encoding mCherry-Actin and V5-hTau1N4R. At 72 h post-infection, cells were processed for immunostaining 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). b Acquisition of TNTs connecting primary neurons and containing endogenous Tau protein. Cells were infected with LVs encoding mCherry-Actin. At 72 h post-infection, neurons were processed for immunostaining analysis using anti-C-Terminal Tau antibodies (Tau-Cter) visualized with an Alexa 488-labeled secondary antibody (green) and anti-acetylated tubulin visualized with an Alexa 647-labeled secondary antibody (white). For (a) and (b), TNTs containing Tau are shown in enlargements (white arrows). c Cells were infected with LVs encoding mCherry-Actin. At 72 h post-infection, neurons were processed for immunocytochemistry using anti-acetylated tubulin visualized with an Alexa 647-labeled secondary antibody (white) and anti C-Terminal Tau antibodies (Tau-Cter) visualized with an Alexa 488-labeled secondary antibody (green). The C-Terminal Tau antibodies used were saturated with Tau proteins for 24 h at 4 °C to block the specific fluorescence signal of tau. Nuclei were labeled with DAPI (blue). For (a), (b) and (c), 1 μM hTau1N4R fibrils were added in the extracellular medium. (d) Cells were infected with LVs encoding mCherry-Actin. TNT formation was activated by ATTO 647-hTau1N4R fibrils. At 72 h later, neurons were processed for immunocytochemistry using anti C-Terminal Tau antibody (Tau-Cter) visualized with an Alexa 488-labeled secondary antibody (green). For (a), (b), (c) and (d), images were acquired using laser-scanning confocal microscopy with a 40× oil-immersion lens (NA 1.3) and processed with ZEN and ImageJ software. Images in (b) and (c) were acquired with the same optical settings and a focal plane was collected. Nuclei were labeled with DAPI (blue). Scale bars: 10 μm

Back to article page