Figure 4
ONAS reduces spontaneous differentiation of neurons during NSC proliferation. (A–I) NSCs were expanded for 1 week as neurospheres and dissociated onto the flat surface or ONAS for 3 additional days in the presence of EGF and FGF2. Then, NSCs were fixed and immunostained with TuJ1 antibody (A–C, green), anti-GFAP antibody (D–F, red) or anti-GalC antibody (G–I, green) to detect neurons, astrocytes or oligodenrocytes, respectively. Nuclei were stained with DAPI (blue). Scale bar, 50 μm. TuJ1-positive (C), GFAP-positive (F) or GalC-positive cells (I) were counted and normalized to total cell number to obtain percentage. Data were presented as mean ± s.e.m. (n = 3). Statistical analysis of data was performed using the Student’s t-test (*P < 0.05).
