Extended Data Fig. 4: Further characterization of blastoid at S4 and the effect of compound from S4 base.

a, b, Immunofluorescent analysis showing the expression of CK8 and GATA6 in blastoids at S4 (a) and E4.5 blastocysts (b). n = 5 S4 blastoids from 2 experiments, n = 3 E4.5 blastocysts from 2 experiments. Scale bars, 20 μm. c, d, Bright field images (c) and formation efficiency (d) of S4 blastoids under different chemical combinations. S4, CHIR-99021 + GA017 + BMP4+bFGF; -C, GA017 + BMP4+bFGF; -G, CHIR-99021 + BMP4+bFGF; -B, CHIR-99021 + GA017+bFGF; -b, CHIR-99021 + GA017 + BMP4. Data are presented as mean ± SD. n = 3, ***p < 0.001, ****p < 0.0001 (One-way ANOVA). p values from left to right: 2.94 × 10−6, 4.14 × 10−4, 1.19 × 10−8, 6.30 × 10−8. Scale bar, 500 μm.