Fig. 3

Comparison of LiPSC- and DF-iPSC-derived neurospheres. a Overview of the culture protocols used in this experiment. b Representative image of floating NSs derived from LiPSCs. Scale bar = 100 μm c NS formation assay using LiPSCs and DF-iPSCs. NSs with a diameter greater than 50 μm were counted (n = 6 independent experiments; means ± SEM; n.s., not significant; Student’s t-test). d–e Morphological analysis of NSs derived from LiPSCs and DF-iPSCs. The circularity and averaged diameter of NSs were quantified (n = 6 independent experiments; means ± SEM; n.s., not significant; Student’s t-test). f–i Expression levels of pluripotent markers (NANOG and OCT4) and neural stem markers (PAX6 and NESTIN) in NSs derived from DF-iPSCs (eKA3, KA11 and KA23) and LiPSCs (LKA10, LKA29, LKA36, LPB1, LPB3 and LPB7). The expression levels were normalized by setting the values from 201B7 [5] to 1.0 (n = 3 independent experiments; means ± SEM). j–m Box-and-whisker plots showing the mRNA transcript levels in NSs derived from DF-iPSCs and LiPSCs. (n = 3 independent experiments; means ± SEM; n.s., not significant; Student’s t-test)