Figure 4

The effect of reduced Npas4 expression on Sox1 expression during neural differentiation of mESCs. (A) The cell proliferation rate during neural differentiation was measured by counting the total number of viable cells at 24 hour intervals. Means and standard deviations of three independent experiments are shown (n = 3). There was no statistically significant difference between the Npas4 knockdown lines and the Empty vector control mESC line (P = 0.2739, non-linear regression). (B) Temporal analysis of Sox1^GFP expression during N2B27 differentiation of Npas4 knockdown and control mESCs as measured by flow cytometry. Means and standard deviations of three independent experiments are shown (n = 3). There was a statistically significant difference between each of the Npas4 knockdown mESC lines and the Empty vector control mESC line (****P ≤0.0001, two-way ANOVA). There was no statistically significant difference between the Empty vector control mESC line and the parental 46C mESC line (P = 0.1898 , two-way ANOVA). (C) Sox1^GFP expression at Day 7 of neural differentiation. Representative flow cytometry histograms are shown for each mESC line. The y-axis shows the number of cells counted while the x-axis shows the GFP fluorescence intensity. The gate used to count GFP⁺ cells is shown. ANOVA, analysis of variance; mESC, mouse embryonic stem cells.