Figure 1.
Shown are the positions of the seven lin-4 binding sites, which are all absent in the n355 mutant allele. In the n355 mutant, the 3′ end of the lin-14 gene is fused to intergenic sequence corresponding to cosmid ZC373. The n536 allele bears a 607-bp deletion in the lin-14 3′UTR, which deletes five of the seven lin-4 binding sites.
Figure 2.
Temporal analyses of lin-14 mRNA, protein, and lin-4 miRNA levels in wild-type and lin-14(n355n679) mutant animals.
(A, C) Quantification of lin-14 mRNA, LIN-14 protein and lin-4 miRNA from early L1 (0 hours post-feeding) to early L2 (24 hours post-feeding) in wild-type (A) and lin-14(n355n679) mutant animals (C). All results are shown relative to the level at the 0 hour time point. Error bars represent SEM for two independent experiments. (B, D) Representative immunoblots showing the abundance of LIN-14 protein in wild-type (B) and lin-14(n355n679) mutant animals (D) over 24 hours of development. Actin serves as a control for the normalization of LIN-14.
Figure 3.
The levels of lin-14 mRNA derived from the wild-type and the lin-14(n536n540) mutant allele bearing a 3′UTR deletion are similar.
Shown are Northern blots of embryos and L4 staged lin-14(n536n540)/szT1 heterozygotes, using probes from the lin-14 (top), histone (middle) and vitellogenin genes (bottom). The levels of 3.5-kb wild-type lin-14 and 2.9-kb lin-14(n536n540) mRNA are almost equal at both the embryonic and L4 stages. Histone mRNA is used as a control for even loading. Vitellogenin mRNA, encoding yolk polypeptides, which is most abundant during oogenesis, is shown to indicate animal stages.