Figure 3
Mature neuronal differentiation and synapse formation between neurons derived from iNPCs.
iNPCs were cultured on collagen IV-coated glass coverslips and subjected to subtype neuronal differentiation. After three weeks differentiation, cells were fixed with 4% PFA and (A) then subjected to double immunostaining with β-tubulin III and MAP-2 (upper panel), β-tubulin III and TH1 (middle panel) and β-tubulin III and GAD67 (low panel). (B) Double immunostaining with β-tubulin III and Synaptophysin antibodies. (a, a′) β-tubulin III (green); (b, b′) Synaptophysin (red); (d, d′) shows a magnification of (c, c′) and arrows indicate the formation of synapses between neurons. Nuclear staining with DAPI (blue). (Scale bars: 50 μm).
