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The differentiation block in I-BET-resistant (I-BETR) diapause-like ES cells. ( A ) Alkaline phosphatase (AP) levels in control and I-BETR ES cells. Scale bar, 100 µm. ( B ) Expression levels of the pluripotency ( left panel) or differentiation-inducing ( right panel) genes in control, 2i-treated, or I-BETR ES cells. RNA expression levels were quantified by qPCR. Values represent normalized mean ± SD. n = 3. ( C ) Colony morphology and/or alkaline phosphatase (AP) levels in control and I-BETR ES cells following vehicle or <t>FGF4</t> triggering. Scale bar, 100 µm. The bar graph represents quantification of relative percentage of pluripotent colonies in different groups. Values represent ± SD. n = 9. (n.s.) No significance, (****) P < 0.0001, one-way ANOVA with Dunnett's multiple comparisons test. ( D ) Expression levels of selected pluripotency ( left ) or differentiation-inducing ( right ) genes in control and I-BETR ES cells treated or not treated with FGF4. Error bars indicate SD. n = 3. (n.s.) No significance, (****) P < 0.0001, one-way ANOVA with Dunnett's multiple comparisons test. ( E ) Withdrawal of I-BET (I-BETW) restores the control ES cell-like pluripotency gene expression pattern in I-BETR ES cells. Values represent gene expression levels normalized to the mean of control samples based on TPM values observed by bulk mRNA RNA-seq analysis . ( F ) Generation of chimeras by I-BETR ES cells. The I-BETR ES cells were incubated in I-BET-free medium for 12–14 h and injected into the C57BL/6J blastocysts. White coat color indicates the chimerism.
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The differentiation block in I-BET-resistant (I-BETR) diapause-like ES cells. ( A ) Alkaline phosphatase (AP) levels in control and I-BETR ES cells. Scale bar, 100 µm. ( B ) Expression levels of the pluripotency ( left panel) or differentiation-inducing ( right panel) genes in control, 2i-treated, or I-BETR ES cells. RNA expression levels were quantified by qPCR. Values represent normalized mean ± SD. n = 3. ( C ) Colony morphology and/or alkaline phosphatase (AP) levels in control and I-BETR ES cells following vehicle or <t>FGF4</t> triggering. Scale bar, 100 µm. The bar graph represents quantification of relative percentage of pluripotent colonies in different groups. Values represent ± SD. n = 9. (n.s.) No significance, (****) P < 0.0001, one-way ANOVA with Dunnett's multiple comparisons test. ( D ) Expression levels of selected pluripotency ( left ) or differentiation-inducing ( right ) genes in control and I-BETR ES cells treated or not treated with FGF4. Error bars indicate SD. n = 3. (n.s.) No significance, (****) P < 0.0001, one-way ANOVA with Dunnett's multiple comparisons test. ( E ) Withdrawal of I-BET (I-BETW) restores the control ES cell-like pluripotency gene expression pattern in I-BETR ES cells. Values represent gene expression levels normalized to the mean of control samples based on TPM values observed by bulk mRNA RNA-seq analysis . ( F ) Generation of chimeras by I-BETR ES cells. The I-BETR ES cells were incubated in I-BET-free medium for 12–14 h and injected into the C57BL/6J blastocysts. White coat color indicates the chimerism.
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The differentiation block in I-BET-resistant (I-BETR) diapause-like ES cells. ( A ) Alkaline phosphatase (AP) levels in control and I-BETR ES cells. Scale bar, 100 µm. ( B ) Expression levels of the pluripotency ( left panel) or differentiation-inducing ( right panel) genes in control, 2i-treated, or I-BETR ES cells. RNA expression levels were quantified by qPCR. Values represent normalized mean ± SD. n = 3. ( C ) Colony morphology and/or alkaline phosphatase (AP) levels in control and I-BETR ES cells following vehicle or <t>FGF4</t> triggering. Scale bar, 100 µm. The bar graph represents quantification of relative percentage of pluripotent colonies in different groups. Values represent ± SD. n = 9. (n.s.) No significance, (****) P < 0.0001, one-way ANOVA with Dunnett's multiple comparisons test. ( D ) Expression levels of selected pluripotency ( left ) or differentiation-inducing ( right ) genes in control and I-BETR ES cells treated or not treated with FGF4. Error bars indicate SD. n = 3. (n.s.) No significance, (****) P < 0.0001, one-way ANOVA with Dunnett's multiple comparisons test. ( E ) Withdrawal of I-BET (I-BETW) restores the control ES cell-like pluripotency gene expression pattern in I-BETR ES cells. Values represent gene expression levels normalized to the mean of control samples based on TPM values observed by bulk mRNA RNA-seq analysis . ( F ) Generation of chimeras by I-BETR ES cells. The I-BETR ES cells were incubated in I-BET-free medium for 12–14 h and injected into the C57BL/6J blastocysts. White coat color indicates the chimerism.
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Image Search Results


The differentiation block in I-BET-resistant (I-BETR) diapause-like ES cells. ( A ) Alkaline phosphatase (AP) levels in control and I-BETR ES cells. Scale bar, 100 µm. ( B ) Expression levels of the pluripotency ( left panel) or differentiation-inducing ( right panel) genes in control, 2i-treated, or I-BETR ES cells. RNA expression levels were quantified by qPCR. Values represent normalized mean ± SD. n = 3. ( C ) Colony morphology and/or alkaline phosphatase (AP) levels in control and I-BETR ES cells following vehicle or FGF4 triggering. Scale bar, 100 µm. The bar graph represents quantification of relative percentage of pluripotent colonies in different groups. Values represent ± SD. n = 9. (n.s.) No significance, (****) P < 0.0001, one-way ANOVA with Dunnett's multiple comparisons test. ( D ) Expression levels of selected pluripotency ( left ) or differentiation-inducing ( right ) genes in control and I-BETR ES cells treated or not treated with FGF4. Error bars indicate SD. n = 3. (n.s.) No significance, (****) P < 0.0001, one-way ANOVA with Dunnett's multiple comparisons test. ( E ) Withdrawal of I-BET (I-BETW) restores the control ES cell-like pluripotency gene expression pattern in I-BETR ES cells. Values represent gene expression levels normalized to the mean of control samples based on TPM values observed by bulk mRNA RNA-seq analysis . ( F ) Generation of chimeras by I-BETR ES cells. The I-BETR ES cells were incubated in I-BET-free medium for 12–14 h and injected into the C57BL/6J blastocysts. White coat color indicates the chimerism.

Journal: Genes & Development

Article Title: Transcriptional derepression of negative regulators of MAP kinase supports maintenance of diapause ES cells in the pluripotent state

doi: 10.1101/gad.353143.125

Figure Lengend Snippet: The differentiation block in I-BET-resistant (I-BETR) diapause-like ES cells. ( A ) Alkaline phosphatase (AP) levels in control and I-BETR ES cells. Scale bar, 100 µm. ( B ) Expression levels of the pluripotency ( left panel) or differentiation-inducing ( right panel) genes in control, 2i-treated, or I-BETR ES cells. RNA expression levels were quantified by qPCR. Values represent normalized mean ± SD. n = 3. ( C ) Colony morphology and/or alkaline phosphatase (AP) levels in control and I-BETR ES cells following vehicle or FGF4 triggering. Scale bar, 100 µm. The bar graph represents quantification of relative percentage of pluripotent colonies in different groups. Values represent ± SD. n = 9. (n.s.) No significance, (****) P < 0.0001, one-way ANOVA with Dunnett's multiple comparisons test. ( D ) Expression levels of selected pluripotency ( left ) or differentiation-inducing ( right ) genes in control and I-BETR ES cells treated or not treated with FGF4. Error bars indicate SD. n = 3. (n.s.) No significance, (****) P < 0.0001, one-way ANOVA with Dunnett's multiple comparisons test. ( E ) Withdrawal of I-BET (I-BETW) restores the control ES cell-like pluripotency gene expression pattern in I-BETR ES cells. Values represent gene expression levels normalized to the mean of control samples based on TPM values observed by bulk mRNA RNA-seq analysis . ( F ) Generation of chimeras by I-BETR ES cells. The I-BETR ES cells were incubated in I-BET-free medium for 12–14 h and injected into the C57BL/6J blastocysts. White coat color indicates the chimerism.

Article Snippet: For FGF4-driven ES cell differentiation, 10 ng/μL recombinant FGF4 (R&D Systems 235-F4) was added on day 0 together with 1 μg/μL heparin (Sigma-Aldrich H3149).

Techniques: Blocking Assay, Control, Expressing, RNA Expression, Gene Expression, RNA Sequencing, Incubation, Injection