Immunofluorescence analysis of ANSCs cultured on various substrates in the presence of 1 µµM retinoic acid and 2% fetal bovine serum for 7 days.

<p>Cell nuclei (blue) were stained using DAPI as a counter staining. Quantification of staining results is shown on (A) 10 µm width substrates of each pattern and (B) 2 µm width substrates of each pattern. Error bars represent mean ± SEM (n = 3, more than 400 cells were counted for each sample). (C) Representative fluorescent images of cells stained positive for (c1) Tuj-1 and nestin and positive for (c2) GFAP and O4. (D) Western blot analysis of nestin, Tuj-1, GFAP and RIP expression in ANSCs on each substrate under differentiation conditions for 7 days. (*<i>p</i><0.05 and **<i>p</i><0.01). Scale bar = 50 µm (C).