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• (B) Survival rate of single and double mutants to indicate the functional relationship between ain-1 and age-1.
• The reporter construct, the control plasmid, and a transformation marker plasmid were coinjected into worms to generate the extrachromosomal arrays for analysis.
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• This result suggests that the high expression of miR-71 during L1 diapause is induced or maintained by other signaling pathways.
• Components of the InsR pathway, including age-1, have recently been predicted to be targets of miR-71 in its role in aging (14).
• If the 3′UTR of age-1 or unc-31 is repressed by miR-71, the GFP expression will be repressed in tissues where miR-71 is expressed in wild-type worms, but derepressed in the same tissues of mir-71(lf) worms.

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Numerous animal species across multiple phyla enter developmental arrest for long-term survival in unfavorable environments and resume development upon stress removal. Such lagged trait recovery, combined with rapid invasive recovery, suggests potential for longer-term shifts in grassland composition and function. Improving social and territorial infrastructure and services, including social protection and welfare systems, the inclusion of disadvantaged groups; supporting employment and skills development; creating high-quality, stable jobs. Explore the pages below to find out about your country’s recovery and resilience plan and how it is being implemented. Starting from its 2022 cycle, the European Semester process was adapted to take into account the creation of the Recovery and Resilience Facility and the implementation of the recovery and resilience plans.
It is also worth mentioning that multiple components of the InsR pathway, including age-1, pdk-1, akt-2, and daf-16, are predicted to be targets of the let-7 family miRNAs. Our data provide the experimental evidence that two components of the InsR pathway are likely direct targets of miR-71 in its role in a specific physiological process, L1 diapause (see a model in Fig. S5). Components of the InsR pathway, including age-1, have recently been predicted to be targets of miR-71 in its role in aging (14).
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(D) A representative chart of the L1 starvation survival rates of different miRNA mutants. However, it remains unclear how, and to what extent, miRNAs coordinate animal survival and development in response to stresses. However, the mechanisms that coordinate the long-term survival, overall developmental arrest, and reinitiation remain to be investigated. However, when newly hatched L1 worms encounter an environment with no food, developmental programs arrest and the worm enters L1 diapause.

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• (C) The poor survival rate of daf-16(mu86, null) was enhanced by mir-71(lf).
• The reporter is strongly expressed in H and V cells in both wild-type and mir-71(lf) worms.
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• Unlike classical heterochronic miRNAs such as lin-4 and let-7, the role of miR-71 in vulval cell division is essential in animals recovering from starvation-induced L1 diapause, but not in animals hatched on plates with food.
• To determine the functional relationship of miR-71 with LIN-42 and LIT-1, mir-71(lf); lin-42(lf) L1 worms were starved for 4 d and recovered on lit-1(RNAi) plates.
• 3′UTRs of genes of interest were cloned into the modified pPD129.57 vector as described previously (18).
• We asked whether the expression of miR-71 was regulated by DAF-16, which is required during L1 diapause for long-term survival (2).

Consistent with the observation described above, the 4-d–starved mir-71(lf) mutants recovering on the RNAi control plates displayed the highly penetrant retarded defect in VPC division. If this were true, the starved mir-71(lf); daf-16(lf) double-mutant worms should show a slow growth phenotype similar to that of daf-16(lf) worms, but no specific VPC timing defect. (H) Fluorescence and DIC images showing that a lin-42 3′UTR reporter was repressed in mir-71(+) worms (2/2 transgenic lines) and prominently derepressed in mir-71(−) worms (2/2 transgenic lines). We thus asked whether miR-71 was required for the reinitiation of developmental programs during the recovery phase after L1 starvation. These results suggest that miR-71 regulates the expression of unc-31 and age-1 through their 3′UTRs. Note that there are extra GFP-positive cells (red arrows) in mir-71(lf) mutants.
To investigate the roles of miRNAs in animal survival during starvation-induced L1 diapause, we impaired the overall miRISC function with loss-of-function (lf) mutants of ain-1 (ku322, ku425, and tm3681) and revery play login ain-2(tm2432) and examined their L1 starvation survival rate (Materials and Methods). The strong suppression of the mir-71(lf) defect by hbl-1(RNAi), and the relatively weak effect of miR-71 on hbl-1 expression, are consistent with the idea that miR-71 exerts its role by modulating activities of multiple genes related to hbl-1 function in developmental timing. In contrast, the nuclear-localized GFP expression under the control of the 3′UTR of age-1(Fig. 3 C and D) or unc-31 (Fig. 3 E and F) was strongly repressed in the control worms, but prominently derepressed in mir-71(lf) mutant worms. If the 3′UTR of age-1 or unc-31 is repressed by miR-71, the GFP expression will be repressed in tissues where miR-71 is expressed in wild-type worms, but derepressed in the same tissues of mir-71(lf) worms. (A) The mir-71(n4115, lf) mutant displayed severe reduction in L1 starvation survival rate, and the reduced survival rate of mir-71(lf) was suppressed by a reduction-of-function allele of age-1(hx546). (C) The reduced L1 starvation survival rate of ain-1(lf) mutants was significantly suppressed by a null allele of unc-31.