Fig. 3

Effect of vitamin C supplementation on PGCLC induction. a Bright field and fluorescence images of day 4 EBs stimulated by no cytokines or by various concentrations of vitamin C (0, 50, 100, 200 μg/ml) in addition to cytokines; n = 5 independent experiments. b FACS analysis of mKate (+) cells induction stimulated by various concentrations vitamin C (0, 50, 100, 200 μg/ml) with or without cytokines; n = 5 independent experiments. c Scatter dot plot for the percentage of mKate (+) cells in day 4 EBs stimulated by different concentrations of vitamin C (0, 50, 100, 200 μg/ml); n = 5 independent experiments. Data are presented as mean ± SD. Statistical analysis was performed by one-way analysis of variance. *p < 0.05. d Scatter dot plot for diameters of day 4 EBs stimulated by different concentrations of vitamin C (0, 50, 100, 200 μg/ml); n = 5 independent experiments. Data are presented as mean ± SD. Statistical analysis was performed by one-way analysis of variance. *p < 0.05. e Scatter dot plot for the percentage of EpCAM and INTEGRINα6 double-positive cells in day 4 EBs stimulated by different concentrations of vitamin C (0, 50, 100, 200 μg/ml); n = 6 independent experiments. Data are presented as mean ± SD. Statistical analysis was performed by one-way analysis of variance. *p < 0.05. f Expression analysis by RT-qPCR for day 4 EBs stimulated by different of concentrations of vitamin C (0, 50, 100, 200 μg/ml). Relative expression levels are shown with normalization to hESCs. n = 3 independent experiments. Data are presented as mean ± SD. Statistical analysis was performed by one-way analysis of variance. *p < 0.05