Fig. 6

Effects of LPS on neurosphere formation capacities during ESC-to-NSC transition. A The different doses of LPS (5, 50, 500 and 5000 ng/mL), supplied through the ESC culture medium, did not affect 46C ES cell viability. Data are presented as means ± SDs (n ≥ 3). Significant difference and P values were analysed by one-way ANOVA with the Tukey’s multiple comparisons test. B Schematic of LPS treatment during ESC-to-NSC transition. Representative images of the effects of LPS treatment on neurosphere formation during ESC-to-NSC transition. Scale bar, 100 µm. Quantification of neurosphere formation after LPS treatment during ESC-to-NSC transition. Data are presented as means ± SEM (n ≥ 3). Significant difference and P values were analysed by one-way ANOVA with the Tukey’s multiple comparisons test. C Schematic of the effects of LPS treatment on self-renewal (neurosphere reformation) capacity of ESC-derived NSCs after treatment with different doses of LPS (5, 50, 500 and 5000 ng/mL). Data are presented as means ± SEM (n ≥ 3). Significant difference and P values were analysed by one-way ANOVA with the Tukey’s multiple comparisons test. Schematic of effects of LPS (500 or 5000 ng/mL) during D ESC-to-NSC transition (days 0–7) and E NSC-to-neural differentiation transition (days 7–14) on neural differentiation capacities of ESCs and ESC-derived NSCs; differentiated cells were assessed on day 14. Effects of LPS on the mRNA expression levels of the neuronal markers Tuj1 and Map2; the astrocytic marker Gfap; the oligodendrocyte markers Oligo2; and inflammatory marker genes Tlr2, Tlr4, IL1b and Tnfa after neural differentiation, as evaluated using quantitative PCR. Data are presented as means ± SEM (n = 3). *P < 0.05, **P < 0.05 by one-way ANOVA with the Tukey’s multiple comparisons test. SFEB, serum-free embryonic body; N2B27, Dulbecco’s modified Eagle medium supplemented with N2 and B27