Figure 2. GSK-3β is predominantly expressed in TGNs. Cultured primary cells were treated with 1.0 mM SNAP for 2 h, and then cultured in NB medium without SNAP for an additional 24 h. After fixing with 4% paraformaldehyde, double immunofluorescent staining was performed using primary antibodies against GSK-3β (red) and Tuj1 (green), followed by counterstaining with Heochst33342 (blue) for nuclei. Scale bar = 20 μm. Arrows indicate representative Tuj1-negative cells.