Figure 2.

In Vitro Differentiation of p53-KO haESCs

(A) Morphology of 7-day haEBs derived from p53 KO-1 cells. Scale bar, 100 μm.

(B) DNA-content analysis of 7-day haEBs derived from p53 KO-1 cells. The percentage of the 1n (G₀/G₁) peak in haEBs was 21.7%.

(C) Expression levels of pluripotency and differentiation genes (Rex1, Hand1, Cdx2, Fgf5, Nestin, and T) in haEBs and haESCs determined by qPCR (n = 3 independent experiments). Data presented as mean ± SEM. t test: ^∗∗∗p < 0.001.

(D) Morphology of haEpiLCs derived from p53 KO-1 cells. Scale bar, 100 μm.

(E) DNA-content analysis of haEpiLCs derived from p53 KO-1 cells. The percentage of the 1n (G₀/G₁) peak in haEpiLCs was 21.8%.

(F) Immunofluorescence of OCT4 and NANOG in haEpiLCs derived from p53 KO-1 cells. DNA was stained with DAPI. Scale bar, 50 μm.

(G) Immunofluorescence of TFE3 in haEpiLCs derived from p53 KO-1 cells and haESCs. DNA was stained with DAPI. Scale bar, 50 μm.

(H) Expression levels of pluripotency genes (Oct4, Nanog, Rex1, and Klf4) in WT-haESCs, WT-EpiSCs, and haEpiLCs determined by qPCR (n = 3 independent experiments). Data presented as mean ± SEM. t test: ^∗∗∗p < 0.001.

(I) Expression levels of EpiSC-specific marker genes (Fgf5 and Otx2) in WT-haESCs, WT-EpiSCs and haEpiLCs determined by qPCR (n = 3 independent experiments). Data presented as mean ± SEM. t test: ^∗∗∗p < 0.001.

(J) Immunostaining of NESTIN and TUJ1 in 10-day differentiation cell cultures of haEpiLCs. Scale bar, 50 μm.

(K) Morphology of haNSCLCs derived from p53 KO-1 cells. Scale bar, 100 μm.

(L) DNA-content analysis of haNSCLCs derived from p53 KO-1 cells. The percentage of the 1n (G₀/G₁) peak in haNSCLCs was 28.5%.

(M) Immunofluorescence of OCT4 and NANOG in haNSCLCs. DNA was stained with DAPI. Scale bar, 50 μm.

(N) Immunofluorescence of neural stem cell-specific markers in haNSCLCs, PAX6, SOX1, and NESTIN. DNA was stained with DAPI. Scale bar, 50 μm.

(O) Immunostaining of oligodendrocytes (O4) and astrocytes (GFAP) derived from p53 KO-1. Scale bar, 50 μm.

(P) Immunostaining of neuronal specific markers (MAP2, NEUN, and TUJ1) in neurons derived from p53 KO-1. Scale bar, 50 μm.

See also Figure S2 and Table S4.