Fig. 2

Dip2b loss-of-function disrupts neural differentiation in the N2B27 medium. A Schematic diagram showing differentiation of mESCs in N2B27 medium. B qRT-PCR of Dip2b expression at days 0, 4, 8 post differentiation (n = 3). C Representative morphological images of WT and KO cells at day 3 post differentiation (n = 3). Scale bar, 250 μm. D Flow cytometric analysis showing the percentage of Sox1-GFP positive cells in differentiated WT and KO cells (n = 3). E Expression of marker genes by qRT-PCR in WT and KO cells at days 4 and 7 post differentiation (n = 3). F and G Immunofluorescence staining of WT and KO cells at days 4 (F) and 10 (G) post differentiation (adherent culture on dishes coated with poly-D-lysine and laminin, n = 3). Scale bars, 50 μm. The error bars represented the mean ± SD, and the significance level was calculated by Student’s t test (two-tailed, equal variance) (ns, not statistically significant; *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001)