Extended Data Fig. 5: Evaluation of developmental relevance of S4 blastoids.
a, Q-PCR analysis showing the effect of LY2955303 on the expression levels of classical totipotency marker genes at S1. Data are presented as mean ± SD, n = 3 biological replicates. **p < 0.01; ****p < 0.0001 (Two-way ANOVA). p values from left to right: 1.75 × 10−5, 9.89 × 10−3, 2.21 × 10−7, 3.73 × 10−6, 1.32 × 10−5, 1.45 × 10−6. b, Representative bright-field images showing the effect of continuous LY2955303 treatment (10 μM, E1.5 to E4.5) on blastocyst formation. Scale bar, 100 μm. Similar images were obtained in at least 3 independent experiments. c, Representative bright-field images showing the effect of continuous LY2955303 treatment (10 μM, S1 to S4) on blastoid formation. Scale bar, 100 μm. d, Histograms showing the effect of continuous LY2955303 treatment (10 μM, S1 to S4) on blastoid formation. Data are presented as mean ± SD, n = 3 biological replicates, ****p < 0.0001 (two-sided Student’s t-test). p = 9.23 × 10−5. e, Representative bright-field images showing the effect of continuous Y27632 treatment (20 μM, E2.5 to E4.5) on blastocyst formation. Scale bar, 100 μm. Similar images were obtained in at least 3 independent experiments. f, Representative bright-field images showing the effect of continuous Y27632 treatment (20 μM, S2 to S4) on blastoid formation. Scale bar, 100 μm. g, Histograms showing the effect of continuous Y27632 treatment (20 μM, S2 to S4) on blastoid formation. Data are presented as mean ± SD, n = 3 biological replicates, ****p < 0.0001 (two-sided Student’s t-test). p = 3.80 × 10−6. h, Representative images showing the formation of decidua in the mouse uterus after S4 blastoids were transferred in vivo. Scale bar, 1 cm (left); 200 μm (right). This experiment was repeated 3 times.
