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. Author manuscript; available in PMC: 2014 Dec 1.
Published in final edited form as: Nat Mater. 2014 Apr 13;13(6):599–604. doi: 10.1038/nmat3945

Figure 1.

Figure 1

Soft substrates promote neuroepithelial conversion while inhibiting neural crest differentiation of hESCs in a BMP4-dependent manner. (a) Schematic diagram showing experimental design of hESC neural induction. hESCs were cultured for 8 d in neural induction medium containing the dual Smad inhibitors, SB 431542 (SB, 10 μM) and LDN 193189 (LDN, 100 nM). (b) Representative immunofluorescence images showing Pax6+ NEs and AP2+ NCs after 8 d of culture with (top; 100 nM) or without (bottom) LDN on vitronectin-coated coverslips and rigid (E = 1,200 kPa) and soft (E = 5 kPa) PMAs. Scale bar, 100 μm. (c) Bar graph showing percentage of Pax6+ NEs on coverslips and rigid and soft PMAs at day 4, 6, and 8. (d) Western blotting showing Pax6 and Sox1 expression levels in hESCs cultured for 8 d on coverslips and rigid and soft PMAs. (e) Bar plot showing percentages of Pax6+ NEs and AP2+ NCs at day 8 as a function of substrate rigidity and LDN concentration. Data represents the mean ± s.e.m with n = 3. P-values were calculated using one-way ANOVA, followed by Tukey post hoc analysis. *, P < 0.05; **, P < 0.01.