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cd1530  (MedChemExpress)


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    MedChemExpress cd1530
    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 + <t>CD1530;</t> 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 .
    Cd1530, supplied by MedChemExpress, used in various techniques. Bioz Stars score: 93/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Images

    1) Product Images from "A continuous totipotent-like cell-based embryo model recapitulates mouse embryogenesis from zygotic genome activation to gastrulation"

    Article Title: A continuous totipotent-like cell-based embryo model recapitulates mouse embryogenesis from zygotic genome activation to gastrulation

    Journal: Nature Cell Biology

    doi: 10.1038/s41556-025-01793-9

    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 .
    Figure Legend 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 .

    Techniques Used: Marker, Expressing

    a , Pseudotime trajectory reconstruction of single cells from subpopulations at S1 and S2. b , Box plots showing the expression of genes enriched in subpopulation 1 at S1 (S1-1) across subpopulations from S1 and S2. Each box depicts the median level along with the first and third quartiles. The whiskers represent the minimum and maximum values. Points indicate outliers, which are defined as values exceeding 1.5 times the interquartile range from the box. The dots in the boxplot represent genes. p value was calculated using the two-sided Wilcoxon rank-sum test, n = 100 (number of genes). p values from left to right: 2.96 × 10 −17 , 3.04 × 10 −22 , 2.16 × 10 −27 , 7.28 × 10 −24 . c , Heatmaps showing the expression of totipotency marker genes in S2 cell aggregates under different chemical combinations. S2, CHIR-99021+Birabresib+CD1530 + 8-Br-cAMP; -C, Birabresib+CD1530 + 8-Br-cAMP; -B, CHIR-99021 + CD1530 + 8-Br-cAMP; -CD, CHIR-99021+Birabresib+8-Br-cAMP; -8Br, CHIR-99021+Birabresib+CD1530. d , Heatmaps showing the expression of Nr5a2 and Tfap2c target genes in S2 cell aggregates under different chemical combinations. S2, CHIR-99021+Birabresib+CD1530 + 8-Br-cAMP; -C, Birabresib+CD1530 + 8-Br-cAMP; -B, CHIR-99021 + CD1530 + 8-Br-cAMP; -CD, CHIR-99021+Birabresib+8-Br-cAMP; -8Br, CHIR-99021+Birabresib+CD1530. e , Expression of marker genes for epiblast, trophectoderm and primitive endoderm in S3 cell aggregates under different chemical combinations. S3, CHIR-99021+Birabresib+CD1530 + 8-Br-cAMP; -C, Birabresib+CD1530 + 8-Br-cAMP; -B, CHIR-99021 + CD1530 + 8-Br-cAMP; -CD, CHIR-99021+Birabresib+8-Br-cAMP; -8Br, CHIR-99021+Birabresib+CD1530.
    Figure Legend Snippet: a , Pseudotime trajectory reconstruction of single cells from subpopulations at S1 and S2. b , Box plots showing the expression of genes enriched in subpopulation 1 at S1 (S1-1) across subpopulations from S1 and S2. Each box depicts the median level along with the first and third quartiles. The whiskers represent the minimum and maximum values. Points indicate outliers, which are defined as values exceeding 1.5 times the interquartile range from the box. The dots in the boxplot represent genes. p value was calculated using the two-sided Wilcoxon rank-sum test, n = 100 (number of genes). p values from left to right: 2.96 × 10 −17 , 3.04 × 10 −22 , 2.16 × 10 −27 , 7.28 × 10 −24 . c , Heatmaps showing the expression of totipotency marker genes in S2 cell aggregates under different chemical combinations. S2, CHIR-99021+Birabresib+CD1530 + 8-Br-cAMP; -C, Birabresib+CD1530 + 8-Br-cAMP; -B, CHIR-99021 + CD1530 + 8-Br-cAMP; -CD, CHIR-99021+Birabresib+8-Br-cAMP; -8Br, CHIR-99021+Birabresib+CD1530. d , Heatmaps showing the expression of Nr5a2 and Tfap2c target genes in S2 cell aggregates under different chemical combinations. S2, CHIR-99021+Birabresib+CD1530 + 8-Br-cAMP; -C, Birabresib+CD1530 + 8-Br-cAMP; -B, CHIR-99021 + CD1530 + 8-Br-cAMP; -CD, CHIR-99021+Birabresib+8-Br-cAMP; -8Br, CHIR-99021+Birabresib+CD1530. e , Expression of marker genes for epiblast, trophectoderm and primitive endoderm in S3 cell aggregates under different chemical combinations. S3, CHIR-99021+Birabresib+CD1530 + 8-Br-cAMP; -C, Birabresib+CD1530 + 8-Br-cAMP; -B, CHIR-99021 + CD1530 + 8-Br-cAMP; -CD, CHIR-99021+Birabresib+8-Br-cAMP; -8Br, CHIR-99021+Birabresib+CD1530.

    Techniques Used: Expressing, Marker



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    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 + <t>CD1530;</t> 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 .
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    Image Search Results


    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 .

    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

    a , Pseudotime trajectory reconstruction of single cells from subpopulations at S1 and S2. b , Box plots showing the expression of genes enriched in subpopulation 1 at S1 (S1-1) across subpopulations from S1 and S2. Each box depicts the median level along with the first and third quartiles. The whiskers represent the minimum and maximum values. Points indicate outliers, which are defined as values exceeding 1.5 times the interquartile range from the box. The dots in the boxplot represent genes. p value was calculated using the two-sided Wilcoxon rank-sum test, n = 100 (number of genes). p values from left to right: 2.96 × 10 −17 , 3.04 × 10 −22 , 2.16 × 10 −27 , 7.28 × 10 −24 . c , Heatmaps showing the expression of totipotency marker genes in S2 cell aggregates under different chemical combinations. S2, CHIR-99021+Birabresib+CD1530 + 8-Br-cAMP; -C, Birabresib+CD1530 + 8-Br-cAMP; -B, CHIR-99021 + CD1530 + 8-Br-cAMP; -CD, CHIR-99021+Birabresib+8-Br-cAMP; -8Br, CHIR-99021+Birabresib+CD1530. d , Heatmaps showing the expression of Nr5a2 and Tfap2c target genes in S2 cell aggregates under different chemical combinations. S2, CHIR-99021+Birabresib+CD1530 + 8-Br-cAMP; -C, Birabresib+CD1530 + 8-Br-cAMP; -B, CHIR-99021 + CD1530 + 8-Br-cAMP; -CD, CHIR-99021+Birabresib+8-Br-cAMP; -8Br, CHIR-99021+Birabresib+CD1530. e , Expression of marker genes for epiblast, trophectoderm and primitive endoderm in S3 cell aggregates under different chemical combinations. S3, CHIR-99021+Birabresib+CD1530 + 8-Br-cAMP; -C, Birabresib+CD1530 + 8-Br-cAMP; -B, CHIR-99021 + CD1530 + 8-Br-cAMP; -CD, CHIR-99021+Birabresib+8-Br-cAMP; -8Br, CHIR-99021+Birabresib+CD1530.

    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 , Pseudotime trajectory reconstruction of single cells from subpopulations at S1 and S2. b , Box plots showing the expression of genes enriched in subpopulation 1 at S1 (S1-1) across subpopulations from S1 and S2. Each box depicts the median level along with the first and third quartiles. The whiskers represent the minimum and maximum values. Points indicate outliers, which are defined as values exceeding 1.5 times the interquartile range from the box. The dots in the boxplot represent genes. p value was calculated using the two-sided Wilcoxon rank-sum test, n = 100 (number of genes). p values from left to right: 2.96 × 10 −17 , 3.04 × 10 −22 , 2.16 × 10 −27 , 7.28 × 10 −24 . c , Heatmaps showing the expression of totipotency marker genes in S2 cell aggregates under different chemical combinations. S2, CHIR-99021+Birabresib+CD1530 + 8-Br-cAMP; -C, Birabresib+CD1530 + 8-Br-cAMP; -B, CHIR-99021 + CD1530 + 8-Br-cAMP; -CD, CHIR-99021+Birabresib+8-Br-cAMP; -8Br, CHIR-99021+Birabresib+CD1530. d , Heatmaps showing the expression of Nr5a2 and Tfap2c target genes in S2 cell aggregates under different chemical combinations. S2, CHIR-99021+Birabresib+CD1530 + 8-Br-cAMP; -C, Birabresib+CD1530 + 8-Br-cAMP; -B, CHIR-99021 + CD1530 + 8-Br-cAMP; -CD, CHIR-99021+Birabresib+8-Br-cAMP; -8Br, CHIR-99021+Birabresib+CD1530. e , Expression of marker genes for epiblast, trophectoderm and primitive endoderm in S3 cell aggregates under different chemical combinations. S3, CHIR-99021+Birabresib+CD1530 + 8-Br-cAMP; -C, Birabresib+CD1530 + 8-Br-cAMP; -B, CHIR-99021 + CD1530 + 8-Br-cAMP; -CD, CHIR-99021+Birabresib+8-Br-cAMP; -8Br, CHIR-99021+Birabresib+CD1530.

    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: Expressing, Marker

    a Western blot analysis showing the levels of histone H3 and H4 acetylation and H3K79me2 of EPS and TPS cells. Similar results were obtained in at least 2 independent experiments. b qPCR analysis of expression levels of classical RAR downstream target genes in EPS and TPS cells. n = 3 biological replicates. c qPCR analysis of expression levels of representative totipotency marker genes on day 3 upon treatment of different small molecule combinations. In the CPEC condition, EPZ004777, VPA, CD1530 and CHIR 99021 were replaced by small molecules targeting DOT1L, HDAC, RA signaling and GSK3β, respectively. n = 2 technical replicates. Similar results were obtained in at least 2 independent experiments. EPS, EPS cells; Basal, EPS cells cultured in the basal medium of CPEC condition. EPZ rep, VPA rep, CD1530 rep and CHIR rep indicate small molecules that target DOT1L, HDAC, RA and GSK3β, respectively. d qPCR analysis of Hdac1 and Hdac2 expression in EPS cells after shRNA knockdown of Hdac1/2 . sh Hdac , Hdac1/2 shRNA. n = 2 biological replicates. e qPCR analysis of expression levels of representative totipotency marker genes on day 3 after knocking down Hdac1/2 in EPS cells during TPS cell induction. EPS, EPS cells; CPEC-V + sh Hdac , replacement of VPA with Hdac1/2 knockdown. n = 2 biological replicates. f qPCR analysis of Dot1l expression in EPS cells after shRNA knockdown of Dot1l . sh Dot1l , Dot1l shRNA. n = 3 biological replicates. g qPCR analysis of expression levels of representative totipotency marker genes on day 3 after knocking down Dot1l in EPS cells during TPS cell induction. EPS, EPS cells; CPEC-E + sh Dot , replacement of EPZ004777 with Dot1l knockdown. n = 3 biological replicates. h qPCR analysis of the effect of inhibiting RAR signaling on totipotency induction (left panel) and maintenance (right panel) in TPS cells. Expression of representative totipotency marker genes were analyzed. RARγi, LY2955303; RARα/βi, LE135; RXRi, UVI3003. n = 3 biological replicates. i qPCR analysis of the effect of CPEC chemical cocktail on totipotency maintenance in early preimplantation embryos. Small molecules were added from the 2-cell embryo stage for 2 days. DMSO was used as a negative control. 2C, 2-cell embryo. n = 2 biological replicates. j qPCR analysis of the effect of Dux knockdown on maintaining totipotency in TPS cells. Expression of representative totipotency marker genes were analyzed. Dux KD, dux knockdown. n = 2 biological replicates. k qPCR analysis of the effect of Dux knockdown on inducing totipotency from EPS cells. Expression of representative totipotency marker genes were analyzed. Dux KD, dux knockdown. n = 2 biological replicates. l qPCR analysis of the effect of p53 knockdown on inducing totipotency from EPS cells. Expression of representative totipotency marker genes were analyzed. p53 KD, p53 knockdown. n = 2 biological replicates.

    Journal: Cell Research

    Article Title: Derivation of totipotent-like stem cells with blastocyst-like structure forming potential

    doi: 10.1038/s41422-022-00668-0

    Figure Lengend Snippet: a Western blot analysis showing the levels of histone H3 and H4 acetylation and H3K79me2 of EPS and TPS cells. Similar results were obtained in at least 2 independent experiments. b qPCR analysis of expression levels of classical RAR downstream target genes in EPS and TPS cells. n = 3 biological replicates. c qPCR analysis of expression levels of representative totipotency marker genes on day 3 upon treatment of different small molecule combinations. In the CPEC condition, EPZ004777, VPA, CD1530 and CHIR 99021 were replaced by small molecules targeting DOT1L, HDAC, RA signaling and GSK3β, respectively. n = 2 technical replicates. Similar results were obtained in at least 2 independent experiments. EPS, EPS cells; Basal, EPS cells cultured in the basal medium of CPEC condition. EPZ rep, VPA rep, CD1530 rep and CHIR rep indicate small molecules that target DOT1L, HDAC, RA and GSK3β, respectively. d qPCR analysis of Hdac1 and Hdac2 expression in EPS cells after shRNA knockdown of Hdac1/2 . sh Hdac , Hdac1/2 shRNA. n = 2 biological replicates. e qPCR analysis of expression levels of representative totipotency marker genes on day 3 after knocking down Hdac1/2 in EPS cells during TPS cell induction. EPS, EPS cells; CPEC-V + sh Hdac , replacement of VPA with Hdac1/2 knockdown. n = 2 biological replicates. f qPCR analysis of Dot1l expression in EPS cells after shRNA knockdown of Dot1l . sh Dot1l , Dot1l shRNA. n = 3 biological replicates. g qPCR analysis of expression levels of representative totipotency marker genes on day 3 after knocking down Dot1l in EPS cells during TPS cell induction. EPS, EPS cells; CPEC-E + sh Dot , replacement of EPZ004777 with Dot1l knockdown. n = 3 biological replicates. h qPCR analysis of the effect of inhibiting RAR signaling on totipotency induction (left panel) and maintenance (right panel) in TPS cells. Expression of representative totipotency marker genes were analyzed. RARγi, LY2955303; RARα/βi, LE135; RXRi, UVI3003. n = 3 biological replicates. i qPCR analysis of the effect of CPEC chemical cocktail on totipotency maintenance in early preimplantation embryos. Small molecules were added from the 2-cell embryo stage for 2 days. DMSO was used as a negative control. 2C, 2-cell embryo. n = 2 biological replicates. j qPCR analysis of the effect of Dux knockdown on maintaining totipotency in TPS cells. Expression of representative totipotency marker genes were analyzed. Dux KD, dux knockdown. n = 2 biological replicates. k qPCR analysis of the effect of Dux knockdown on inducing totipotency from EPS cells. Expression of representative totipotency marker genes were analyzed. Dux KD, dux knockdown. n = 2 biological replicates. l qPCR analysis of the effect of p53 knockdown on inducing totipotency from EPS cells. Expression of representative totipotency marker genes were analyzed. p53 KD, p53 knockdown. n = 2 biological replicates.

    Article Snippet: Small molecules replacing CD1530 included AM580 (0.05 μM; Selleck, S2933), ch55 (0.05 μM; MCE, HY-107397), Palovarotene (0.01 μM; MCE, HY-14799), CD3254 (50 nM; MCE, CD3254) and RA (0.5 μM; MCE, HY-14649).

    Techniques: Western Blot, Expressing, Marker, Cell Culture, shRNA, Knockdown, Negative Control