Fig. 6

Tripartite synapse structure in the co-culture system and the cell-cell interaction between the induced neurons and astrocytes. a Immunocytochemistry image showing the neuronal synaptic markers (VGLUT2 and PSD95) and the astrocyte marker (GFAP). Scale bar; 10 μm. b Immuno-electron microscopy (iEM) image showing the neuronal synaptic terminals, and astrocytic process stained with S100β. Scale bar; 200 nm. c Evaluation of neuronal excitability measured by Ca imaging using Fluo-8 indicator in the co-culture model after treatment with 20 μM TFB-TBOA (Excitatory Amino Acid Transporter (EAAT) 1,2 inhibitor). d Representative waveforms of neuronal activity (Ca²⁺ oscillations) of the co-culture model before and after treatment with 20 μM TFB-TBOA. e Quantification of neuronal firing (Spikes/Min and ΔF/F₀) of the co-culture model after treatment with 20 μM TFB-TBOA (n = 30) compared to the treatment with DMSO (n = 30) (three independent experiments). ****p < 0.0001 (Mann-Whitney test)