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Fig. 4 | Biological Procedures Online

Fig. 4

From: A 3D human co-culture to model neuron-astrocyte interactions in tauopathies

Fig. 4

Seed-independent intraneuronal tau aggregation. Neural progenitors were transduced with mCherry-tagged FTDtau1 or EGFP-tagged FTDtau1+2, mixed separately (A, B, D) or combined (C) with human primary astrocytes and tau pathology was assessed at week 4. A 12 µm-thick maximum intensity confocal image projections of 3D-cocultures expressing EGFP-tagged FTDtau1 or FTDtau1+2 (green), immunostained for neurons (MAP2, blue) and a pathological conformation of tau (MC1, magenta). Single channels are shown in greyscale. A zoom of the boxed area indicated in the MC1 channel is shown as merged image and shows overlap of FTDtau1+2 and MC1 within a MAP2-positive neuron. B 11 µm-thick maximum intensity confocal image projection of 3D co-cultures expressing EGFP-tagged FTDtau1 or FTDtau1+2, immunostained for a pathological conformation of tau (MC1), co-labelled for stacked beta-sheets (HS169). Nuclei were visualized using DAPI (blue) and are merged with FTDtau (grey). The HS169 and MC1 single channels are shown in greyscale and the merged image shows that only FTDtau1+2 (blue) overlaps with MC1 (magenta) and HS169 (green). C 10 µm-thick maximum intensity confocal image of a 3D neuron/astrocyte co-culture with neurons expressing either mCherry-tagged FTDtau1 (blue) or EGFP-tagged FTDtau1+2 (green), immunostained for a pathological conformation of tau (MC1, magenta). A zoom of the boxed area indicated in the MC1 channel is shown as merged image and shows that only FTDtau1+2 overlaps with MC1. D 50 µm-thick maximum intensity confocal image projections of a 3D-coculture expressing EGFP-tagged FTDtau1+2 (green), immunostained for neurons (β-3-tubulin, blue) and astrocytes (GFAP, magenta). Single channels are shown in greyscale. A zoom of the boxed area indicated in the GFAP channel is shown as merged image. The arrows in the orthogonal views show an astrocytic extension in contact with a FTDtau1+2-containing neuron

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