Journal: bioRxiv

Article Title: Basal activation of astrocytic Nrf2 in neuronal culture media: challenges and implications for neuron-astrocyte modelling

doi: 10.1101/2024.09.18.613665

Figure Lengend Snippet: (A) LUHMES neurons were seeded onto a layer of primary human astrocytes at day two of the co-culture protocol, then fixed and immunostained at day six post-differentiation for β-III tubulin (green) and the astrocyte-specific marker ALDH1L1 (red). Nuclei were labelled by DNA staining with DAPI (blue). (I) Neurons alone (II) Human astrocytes alone. (II) Nuclei alone. (IV) Merged image of A-C. (V) No antibody, negative control. (VI) Cells immunostained with goat anti-mouse 488 and goat anti-rabbit 594 secondary antibodies alone. Images were captured using a ZEISS LSM 800 confocal microscope with 63x oil lens. Scale bars, 10 µm (B) Effects of Nrf2 activators on NQO1 activity in LUHMES neuron -astrocyte co-cultures maintained in LUHMES differentiation media, following treatment with 18e (10 µM), DMF (40 µM), CDDO-Me (10 nM), and 0.1% DMSO vehicle control, for 25-32hr. (n = 6, three independent experiments). Values derived from each independent vial of cells are denoted by colour and shape.

Article Snippet: Cell nuclei were stained with ProLong TM Diamond Antifade Mountant with DAPI (Thermo Fisher Scientific, P36966).

Techniques: Co-Culture Assay, Marker, Staining, Negative Control, Microscopy, Activity Assay, Control, Derivative Assay