Fig. 2

a Following LDN193189 restriction for 24 h, when cells were cultured under N2, N2 and LDN193189, or dual inhibition medium, the percentage of Nestin⁺ cells increased from 84.55 ± 1.14 % to 96.84 ± 1.33 %. By contrast, no obvious changes were observed for Pax6⁺, SOX1⁺ and SOX2⁺ cells. b Compared with hESCs, OCT4 expression was downregulated significantly only after LDN193189 restriction followed by dual inhibition. NANOG was downregulated significantly after LDN193189 restriction followed by N2, N2 and LDN193189 or dual inhibition medium. c After LDN193189 restriction, when cells were cultured in dual inhibition medium for 5 days, Pax6 expression increased significantly compared with undifferentiated hESCs, culture in N2 supplemented medium or LDN193189 inhibition for 5 days. d After lineage restriction with LDN193189 treatment, 5 days of dual inhibition resulted in significantly higher levels of neural gene expression comparing with the other treatment groups. e After dual lineage restriction, 5 days of continuous LDN1931898 inhibition or dual inhibition resulted in a significantly lower percentage of SOX1⁺ cells. n = 3 for each group