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Elegans Genetic Center (reference 257) and an N2 strain from the laboratory stock, respectively. Wild-type strains A and B are an N2 strain recently obtained from the C. (A) Survival rate curves of wild-type and mutant strains, as indicated. This is consistent with the previous reports that AIN-1 and AIN-2 are functional homologs with overlapping biochemical roles (16, 17). The roles of InsRs have also been implicated in arresting the cell cycle in germ cells and a portion of somatic cells during L1 diapause (2, 4). Contributed new reagents/analytic tools; X.Z., R.Z., and M.H.

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Furthermore, worms that are long-lived due to dietary restriction or decreased mitochondrial respiratory rates are short-lived during L1 diapause, suggesting that the mechanisms controlling L1 starvation survival are different at least in some aspects from those controlling aging (3). We provide evidence that miRNA miR-71 is not required for the animals’ entry into L1 diapause, but plays a critical role in long-term survival by repressing the expression of insulin receptor/PI3K pathway genes and genes acting downstream or in parallel to the pathway. Full device encrypted backups to iTunes will back up both the account listings and private key pairs, but can only be restored on the SAME phone that created the backup. If you enabled third-party account backup, you can recover your accounts on your iOS or Android device. If your organization hasn’t enabled self-service device management, contact your IT Help Desk or Duo service administrator for assistance reactivating the account.

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The RRF funding amounts shown for measures are based on the initial cost estimates provided by the Member States included in the recovery and resilience plans and may as such differ from finalized Commission financial reports, which follow eventual project implementation. The funding amounts shown reflect the initial cost estimates included in the national recovery and resilience plans. Member States use the funds provided by the Recovery and Resilience Facility to implement ambitious reforms and investment to make their economies and societies more sustainable, resilient and prepared for the green and digital transitions. The Scoreboard gives an overview of progress in implementing the Facility and the national recovery and resilience plans.
On the other hand, the role of a particular miRNA (miR-71) is executed by repressing the expression of many genes in multiple pathways. On one hand, we showed that deletions of a good number of miRNAs have varying impacts on the L1 diapause survival rate, although they may effect the rate through different mechanisms. Instead, many specific physiological functions, such as the starvation-induced stress response, are regulated by a miRNA-target network, often involving multiple miRNAs and a large number of their targets. We found that the known developmental timing genes, hbl-1, lin-42, and lit-1, were at the top of the list (TargetScan). To understand how miR-71 affects VPC division, we searched its predicted targets for potential genes involved in regulating developmental timing. These results indicate that miR-71 plays a significant role in larval development of animals recovering from L1 diapause and likely does so by regulating the expression of components of the insulin receptor/DAF-16 pathway, as well as factors acting downstream, or in parallel to, DAF-16.
This will be followed by an ‘ex post evaluation’ in 2028, once the measures included in the recovery plans are fully implemented. The RRF Regulation requires that the Commission provides the European Parliament, the Council, the European Economic and Social Committee and the Committee of the Regions with a mid-term evaluation on the implementation of the Recovery and Resilience Facility. Member States can also amend their plan if they can demonstrate that objective circumstances render the implementation of certain milestones and targets unfeasible. The RRF is also crucial for implementing the REPowerEU plan – the Commission’s response to the socio-economic hardships and global energy market disruption caused by Russia’s invasion of Ukraine.
Compromising overall miRNA function dramatically reduces the survival rate of L1 worms in starvation-induced diapause, and the effect can be significantly suppressed by an age-1/PI3K mutation. Furthermore, miR-71 plays a prominent role in developmental recovery from L1 diapause partly through repressing the expression of certain heterochronic genes. When you restore a backup that contains third-party account information you must enter the recovery password to decrypt the backup. If you opt-in to third-party account backup and restore, and have set an account recovery password, then the app backups to Google Drive (Android) or iCloud (iOS) do include the private key information for your third-party accounts.
To test the hypothesis that these developmental timing genes mediate the regulatory role of miR-71 in larval development during recovery from starvation-induced L1 diapause, we examined whether knocking down HBL-1 function can suppress the retarded VPC timing defect of mir-71(lf). Reduction-of-function mutation (rf) in the age-1/PI3 kinase gene, age-1(hx546), made worms long-lived in the L1 starvation assay and was able to suppress the reduced L1 survival rate of mir-71(lf); the rate of the double mutants was comparable to that of wild type (Fig. 2A). Our genetic analysis indicated that for both L1 diapause survival and developmental recovery functions, miR-71 regulates expressions of genes in both the insulin receptor-dependent and -independent pathways.
Non-Unc stable transgenic lines were maintained, and the expression of GFP and mCherry were observed under a Zeiss Axiovision II microscope. Three days later, the number of worms that were L2 or older was recorded as number of survived worms (Ns), and the survival rate was calculated as Ns/Np, which is an estimation of survived worms in the whole population. MT12993 mir-71(n4115) worms were outcrossed with N2 for four generations before any test except the initial screen.

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• These results indicate that miR-71 plays a significant role in larval development of animals recovering from L1 diapause and likely does so by regulating the expression of components of the insulin receptor/DAF-16 pathway, as well as factors acting downstream, or in parallel to, DAF-16.
• This is consistent with the previous reports that AIN-1 and AIN-2 are functional homologs with overlapping biochemical roles (16, 17).
• (B) Bar graph showing the correlation between the severity of the retarded vulval precursor cell (VPC) timing defect of mir-71(lf) mutants and the duration of L1 starvation.
• The coordinated entrance into developmental arrest, long-term survival, and the reinitiation of development upon food availability are important biological processes to investigate.
• If you can’t open Duo Mobile on your old device, for example, if your phone was lost or damaged, contact your Duo administrator to discuss your account recovery options.
• This result is consistent with the observation that miR-71 is specifically required for the starvation-induced stress response (Fig. S5).
• Note that the daf-16(lf) worms recovering from 3 d of L1 starvation displayed a ∼12-h delay in overall development and that the mir-71(lf); daf-16(lf) double mutants displayed an ∼24-h delay.

If you haven’t enabled third-party account restore in Duo Mobile then app backups to Google account backup (Android) or iCloud (iOS) accounts DO NOT contain any private key or other sensitive data. Duo Mobile’s restore functionality lets you back up Duo-protected accounts and third-party OTP accounts (such as Google or Facebook) for recovery to the same device or to a new device. We speculate that the expression of heterochronic genes controlling the L2/L3 programs, including that of hbl-1 and lin-42, are increased during L1 diapause to arrest the developmental progression, and miR-71 is probably required to suppress these “excess” signals during the recovery phase (Fig. S5). Furthermore, the observed derepression of individual genes by mir-71(lf) seemed too weak to account for the phenotype, consistent with the idea that a prominent phenotype of an miRNA mutation is caused by the collective effect of changing expression in many genes, an important property of miRNA-mediated gene regulation. (F) Fluorescence and DIC images showing that an hbl-1 3′UTR reporter was repressed in mir-71(+) worms and slightly derepressed in mir-71(lf) mutants.

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{(E) DIC images showing that hbl-1(RNAi) caused precocious VPC divisions in late L2/early L3 in both wild-type and mir-71(lf) worms recovered from 4 d of L1 starvation. Note that the daf-16(lf) worms recovering from 3 d of L1 starvation displayed a ∼12-h delay in overall development and that the mir-71(lf); daf-16(lf) double mutants displayed an ∼24-h delay. (C) Bar graph showing that the delayed VPC timing defects of mir-71(lf) worms was suppressed by an unc-31(lf) mutation and partially suppressed by an age-1(rf) mutation. In worms that recovered from 4 d of L1 starvation, we also found that a significant portion of the mir-71(lf) mutants displayed egg-laying defects and overproliferating or precociously reflexed gonads.}