The level of
nhr-91 transcripts was increased in starved
mir-235(
n4504) L1 larvae, compared with wild type. In fed wild-type animals, significant upregulation of
nhr-91 transcripts temporally coincides with downregulation of mature miR-235 after 8 and 10 h of feeding. Similarly, a reporter gene, in which the gfp-pest complimentary DNA was inserted between the promoter and the 3'UTR of
nhr-91 (Pnhr-91::gfp- pest::
nhr-91 3' UTR), also showed a burst of expression in the hypodermis, where Pmir-235::gfp was also expressed, after 8 to 10h of feeding. The level of
nhr-91 transcripts was further increased in
mir-235 mutants compared with wild-type animals after 8 h of feeding, suggesting that
mir-235 functions not only to suppress expression of
nhr-91 during L1 diapause, but also to limit the kinetics of its upregulation when feeding.