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You have provided additional information to address many of the concerns raised in the first round of review, as both referees acknowledge, and your reply to the referees was considered and clear. Martin Pera, University of Melbourne, Australia. Salt Lake City, Utah. We further delineate a genetic pathway downstream of Eprn involving known players in post-transcriptional and epigenetic regulation, specifically Lin28, let-7 and de novo DNA methyltransferases. Include parent Tweet. To clarify the significance of Eprn in the regulation of pluripotency, it would be helpful to provide more information on the knockdown cells. I like this shot from Friday.
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Does Eprn associate with it or not? To assess the effect of Eprn depletion functionally, we conducted colony forming assays.
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We used let-7g as a representative member since all apart from let-7e share the same seed sequence Figure 4—figure supplement 1B. Concomitant knockdown of Nanog and Lin28a abolished this effect Figure 3C. We now demonstrate that Eprn KO cells have the capacity to undergo neuronal and mesendodermal lineage specification.
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|Non-coding transcripts harbouring TE sequences are enriched in ESCs and early embryo development in both mouse and human Fort et al.
Learn more. Primary antibody was carefully washed away with 0.
One explanation for anti-correlated expression could be direct negative regulation of Lin28a by Nanog. Reviewer 2 notes: 1 "There is really no insight into how the lncRNA functions.
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Our study aimed to pinpoint a lncRNA that has a measurable biological function and to reveal the downstream genetic interactions.
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Therefore, the effect of a given miRNA on an individual target has to be investigated experimentally. Two siRNA combination were used per transfection for each target gene knockdown.
Furthermore, we did not observe Lin28a upregulation in Nanog knockdown cells Figure 3—figure supplement 2H. We have modified the text to be more explicit about our goals and to qualify our conclusions. Try again or visit Twitter Status for more information. This is test 1 of 2 to be fully certified.
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|Indeed, no significant enrichment genome-wide was observed in wild type compared to Eprn KO cells Figure 3—figure supplement 2E.
Add this Tweet to your website by copying the code below. By 72 hr, however, GFP expression was fully extinguished from either starting condition Figure 2—figure supplement 2A.
We further delineate a genetic pathway downstream of Eprn involving known players in post-transcriptional and epigenetic regulation, specifically Lin28, let-7 and de novo DNA methyltransferases.
We generated Eprn knockout KO ESCs via sequential gene targeting Figure 2—figure supplement 1 and examined the phenotype in each condition.
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