Fig. 6

Neural differentiation of mESCs by SrC switch-controlled expression of BRN4. A Structure of the SeV(Csy4/RS-Brn4/RS-EGFP) vector. B Morphology of mESCs harboring SeV(Csy4/RS-Brn4/RS-EGFP). SeV(Csy4/RS-Brn4/RS-EGFP)-infected EB5 cells were selected with G418 in the presence or absence of 300 nM Shield1. Morphology and EGFP expression were observed 5 days after infection. Scale bars, 100 μm. C SSEA1 and Nestin expression in mESCs. SeV(Csy4/RS-Brn4/RS-EGFP)-infected EB5 cells were prepared as described in B. SSEA1 and Nestin were immunostained 7 days after the vector infection. Scale bars, 50 μm. D Morphology of differentiated NSCs. SeV(Csy4/RS-Brn4/RS-EGFP)-infected EB5 cells were differentiated into NSCs using NSC medium. Shield1 concentration was changed according to the indicated scheme. Morphology of the cells were observed at days 3, 8, and 13. Scale bars, 100 μm. E Nestin expression in differentiated NSCs. SeV(Csy4/RS-Brn4/RS-EGFP)-infected EB5 cells were differentiated into NSCs as D. Nestin was immunostained at day 7 of the NSC differentiation. Scale bars, 100 μm. F NSC marker expression in differentiated NSCs. SeV(Csy4/RS-Brn4/RS-EGFP)-infected EB5 cells were differentiated into NSCs as described in D. Pax6 and Sox11 mRNA levels in the cells were determined at day 15 of the NSC differentiation. Data are represented as the means ± SEM of three independent experiments. **p < 0.01. G GFAP expression in differentiated astrocytes. NSCs differentiated from SeV(Csy4/RS-Brn4/RS-EGFP)-infected EB5 cells were cultured under the indicated condition before and after astrocyte differentiation. GFAP was immunostained at day 6 of the astrocyte differentiation. Scale bars, 100 μm