Journal: Nature Cell Biology
Article Title: A continuous totipotent-like cell-based embryo model recapitulates mouse embryogenesis from zygotic genome activation to gastrulation
doi: 10.1038/s41556-025-01793-9
Figure Lengend Snippet: a , Schematic of the strategy used for identifying chemical cocktails that supports generation of transiently induced totipotent-like cells. Td, tdTomato. b , Results of the primary chemical screen using mouse EPSCs carrying the MuERV-L -mClover3 reporter. The dashed line indicates the threshold, which is set to two-fold changes. The values were normalized based on log2. n = 3 biological replicates. c , Heatmaps showing the upregulation of totipotent marker genes in mouse EPSCs under different chemical combinations. Samples were treated for three days before their collection. C, CHIR-99021; CE, CHIR-99021+Elvitegravir; CP, CHIR-99021 + PD0325901; CCD, CHIR-99021 + CD1530; S1, CD1530 + PD0325901+Elvitegravir+CHIR-99021. d , Heatmaps showing the upregulation of primitive endoderm genes in mouse EPSCs under different chemical combinations. Samples were treated for three days before their collection. C, CHIR-99021; CCD, CHIR-99021 + CD1530. e , Q-PCR analysis showing the effect of PD0325901 on the expression levels of classical totipotent marker genes. Samples were treated for three days before their collection. CCD, CHIR-99021 + CD1530; +PD, CHIR-99021 + CD1530 + PD0325901. Data are presented as mean ± SEM, n = 3 biological replicates. * p < 0.05; ** p < 0.01; **** p < 0.0001; ns, non-significant (Two-way ANOVA). p values from left to right: 3.31 × 10 −7 , 6.67 × 10 −8 , 1.64 × 10 −7 , 2.04 × 10 −7 , 1.40 × 10 −3 , 7.98 × 10 −5 , 1.89 × 10 −2 . f , Q-PCR analysis showing the effect of PD0325901 on the expression levels of classical primitive endoderm genes. Samples were treated for three days before their collection. CCD, CHIR-99021 + CD1530; +PD, CHIR-99021 + CD1530 + PD0325901. Data are presented as mean ± SEM, n = 3 biological replicates. *** p < 0.001; **** p < 0.0001; ns, non-significant (Two-way ANOVA). p values from left to right: 3.29 × 10 −5 , 8.05 × 10 −11 , 4.60 × 10 −14 , 8.70 × 10 −4 , 0.117. g , Cell doubling times of CCD, +PD, +PD + E(S1), EPSCs and TPSC. CCD, CHIR-99021 + CD1530; +PD, CHIR-99021 + CD1530 + PD0325901; +PD + E (S1), CHIR-99021 + CD1530 + PD0325901+Elvitegravir. Data are presented as mean ± SD, n = 3 biological replicates. **** p < 0.0001 (One-way ANOVA). p values from left to right: 3.35 × 10 −5 , 1.85 × 10 −6 , 3.10 × 10 −11 .
Article Snippet: The totipotency medium consisted of 1640 basal medium supplemented with elvitegravir (1 μM, MCE, HY-14740), CHIR-99021 (3 μM, Selleck, S1263), CD1530 (0.2 μM, MCE, HY-108527) and PD0325901(0.5 μM, Selleck, S1036).
Techniques: Marker, Expressing