Figure 2.

Cellular Localization of OLIG2 in VFOs and DFOs

(A and B) Representatives (A) and quantification (B) of GFP⁺, OLIG2⁺, and GFP⁺/OLIG2⁺ cells in the VFOs and DFOs at week 5 (n = 4 organoids from two hPSC lines). Scale bars, 100 μm in the original images and 50 μm in the enlarged images.

(C) Quantification of the percentage of GFP⁺ cells with nuclear or cytoplasmic OLIG2 expression among total GFP⁺ cells (n = 3 organoids from two hPSC lines). Student's t test: ^∗∗∗p < 0.001.

(D) Western blot showing the cellular localization of OLIG2 extracted from week-5 organoids. The experiments were repeated three times. For each experiment, at least ten VFOs or DFOs derived from OLIG2-GFP hESCs and hiPSCs were pooled for protein extraction and fractionation.

(E) Representative images of NKX2.1⁺/GFP⁺ cells in VFOs (top panels) and PAX6⁺/GFP⁺ cells in DFO (bottom panels). Scale bars, 50 μm in the original images and 10 μm in the enlarged images.

(F) Quantification of percentage of GFP⁺/PAX6⁻ cells and GFP⁺/PAX6⁺ cells in week-5 DFOs (n = 5 organoids from two hPSC lines).