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Esrrb is a direct Nanog target gene that can substitute for Nanog function in pluripotent cells.

Festuccia N, Osorno R, Halbritter F, Karwacki-Neisius V, Navarro P, Colby D, Wong F, Yates A, Tomlinson SR, Chambers I - Cell Stem Cell (2012)

Bottom Line: Moreover, Esrrb can reprogram Nanog(-/-) EpiSCs and can rescue stalled reprogramming in Nanog(-/-) pre-iPSCs.Finally, Esrrb deletion abolishes the defining ability of Nanog to confer LIF-independent ESC self-renewal.These findings are consistent with the functional placement of Esrrb downstream of Nanog.

View Article: PubMed Central - PubMed

Affiliation: MRC Centre for Regenerative Medicine, Institute for Stem Cell Research, School of Biological Sciences, University of Edinburgh, 5 Little France Drive, Edinburgh EH16 4UU, Scotland.

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Loss of Esrrb Impairs Nanog-Driven LIF Independence(A) Schematic representation of the genetic manipulations used to make conditional knockout (Esrrbf/fn) ESCs that have two floxed Esrrb alleles and express Cre-ERT2.(B) Morphology and expression of Oct4 and Esrrb in Esrrbf/fn and deleted EsrrbΔ/Δ lines.(C) Colony formation after clonal density plating and 7 days culture (+/− LIF; values are the average of six independent clones for each indicated line). Error bars: standard deviation of the results obtained from six clones each analyzed in triplicate.(D) Representative morphologies of colonies formed by the indicated lines after 7 days of culture (+/− LIF).See also Figure S7.
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fig7: Loss of Esrrb Impairs Nanog-Driven LIF Independence(A) Schematic representation of the genetic manipulations used to make conditional knockout (Esrrbf/fn) ESCs that have two floxed Esrrb alleles and express Cre-ERT2.(B) Morphology and expression of Oct4 and Esrrb in Esrrbf/fn and deleted EsrrbΔ/Δ lines.(C) Colony formation after clonal density plating and 7 days culture (+/− LIF; values are the average of six independent clones for each indicated line). Error bars: standard deviation of the results obtained from six clones each analyzed in triplicate.(D) Representative morphologies of colonies formed by the indicated lines after 7 days of culture (+/− LIF).See also Figure S7.

Mentions: To determine the requirement of Esrrb in ESC self-renewal, cells homozygous for a conditional Esrrb knockout allele (Esrrbf/fn) (Chen and Nathans, 2007) and expressing Cre-ERT2 were generated (Figures 7A, S7A, and S7B). Tamoxifen treatment of Esrrbf/fn cells increases the degree of differentiation in these cultures. Nonetheless, stable EsrrbΔ/Δ cell lines genetically devoid of Esrrb were readily isolated (Figure S7C and S7D). Although they show an impaired ability to self-renew in clonal assays (Figure 7C), EsrrbΔ/Δ cells can be propagated in FCS/LIF/GMEMβ and maintain Oct4 expression (Figure 7B). These results establish the fact that despite having a clear stimulatory effect on the efficiency of colony formation, Esrrb is formally dispensable for ESC self-renewal.


Esrrb is a direct Nanog target gene that can substitute for Nanog function in pluripotent cells.

Festuccia N, Osorno R, Halbritter F, Karwacki-Neisius V, Navarro P, Colby D, Wong F, Yates A, Tomlinson SR, Chambers I - Cell Stem Cell (2012)

Loss of Esrrb Impairs Nanog-Driven LIF Independence(A) Schematic representation of the genetic manipulations used to make conditional knockout (Esrrbf/fn) ESCs that have two floxed Esrrb alleles and express Cre-ERT2.(B) Morphology and expression of Oct4 and Esrrb in Esrrbf/fn and deleted EsrrbΔ/Δ lines.(C) Colony formation after clonal density plating and 7 days culture (+/− LIF; values are the average of six independent clones for each indicated line). Error bars: standard deviation of the results obtained from six clones each analyzed in triplicate.(D) Representative morphologies of colonies formed by the indicated lines after 7 days of culture (+/− LIF).See also Figure S7.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3473361&req=5

fig7: Loss of Esrrb Impairs Nanog-Driven LIF Independence(A) Schematic representation of the genetic manipulations used to make conditional knockout (Esrrbf/fn) ESCs that have two floxed Esrrb alleles and express Cre-ERT2.(B) Morphology and expression of Oct4 and Esrrb in Esrrbf/fn and deleted EsrrbΔ/Δ lines.(C) Colony formation after clonal density plating and 7 days culture (+/− LIF; values are the average of six independent clones for each indicated line). Error bars: standard deviation of the results obtained from six clones each analyzed in triplicate.(D) Representative morphologies of colonies formed by the indicated lines after 7 days of culture (+/− LIF).See also Figure S7.
Mentions: To determine the requirement of Esrrb in ESC self-renewal, cells homozygous for a conditional Esrrb knockout allele (Esrrbf/fn) (Chen and Nathans, 2007) and expressing Cre-ERT2 were generated (Figures 7A, S7A, and S7B). Tamoxifen treatment of Esrrbf/fn cells increases the degree of differentiation in these cultures. Nonetheless, stable EsrrbΔ/Δ cell lines genetically devoid of Esrrb were readily isolated (Figure S7C and S7D). Although they show an impaired ability to self-renew in clonal assays (Figure 7C), EsrrbΔ/Δ cells can be propagated in FCS/LIF/GMEMβ and maintain Oct4 expression (Figure 7B). These results establish the fact that despite having a clear stimulatory effect on the efficiency of colony formation, Esrrb is formally dispensable for ESC self-renewal.

Bottom Line: Moreover, Esrrb can reprogram Nanog(-/-) EpiSCs and can rescue stalled reprogramming in Nanog(-/-) pre-iPSCs.Finally, Esrrb deletion abolishes the defining ability of Nanog to confer LIF-independent ESC self-renewal.These findings are consistent with the functional placement of Esrrb downstream of Nanog.

View Article: PubMed Central - PubMed

Affiliation: MRC Centre for Regenerative Medicine, Institute for Stem Cell Research, School of Biological Sciences, University of Edinburgh, 5 Little France Drive, Edinburgh EH16 4UU, Scotland.

Show MeSH
Related in: MedlinePlus