Figure 3

Effect of SEMA3A supplementation on axonal growth of hESC- and hiPSC-derived RGCs. (a) Axonal growth of hESC- and hiPSC-derived RGCs is observed by immunostaining of NFL. Axonal growth of hESC- and hiPSC-derived RGCs is starkly inhibited by 100 and 200 ng/ml SEMA3A supplementation from D27−30 (centre and right panels, respectively). The assessment from D27 is performed under RMM supplemented with 1.0% FBS and 100 ng/ml BDNF. (b) Real-time PCR analysis of mRNA expression of axonal markers, TAU, NFL, and TUJ1. In both hESC- and hiPSC-derived RGCs, the expression of, TAU, NFL, and TUJ1 appears to decrease in a dose-dependent manner. Expression levels of all markers are significantly decreased at 200 ng/ml SEMA3A supplementation (ANOVA, Tukey-HSD test, p ≤ 0.05). Scale bar, 100 μm. Error bars indicate±SD. Each column shows an average value for the studied samples. The sample size for all mRNA data is five (n = 5). NS, not significant.