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. 2009 Mar;15(3):400–405. doi: 10.1261/rna.1258309

FIGURE 1.

FIGURE 1.

lin-14 mRNA and protein are down-regulated by lin-4 miRNA. (A) Total RNA samples from N2 wild-type (WT) staged larvae developing on food for the indicated hours were probed for lin-4 miRNA and control 5.8S rRNA by Northern blot analysis. Quality of the RNA samples was assessed by ethidium bromide staining of the small ribosomal RNAs. Mature lin-4 miRNA levels were normalized to 5.8S rRNA and values relative to the 18-h time point are indicated in the black box. (B) The WT RNA used in (A) as well as RNA collected from lin-4(e912) [lin-4(−)] and lin-14(n355) [lin-14(gf)] strains staged for the same time course were subjected to Northern blotting to detect lin-14 and control actin mRNA and 18S rRNA. Quality of the RNA samples was assessed by ethidium bromide staining of the large ribosomal RNAs. The lin-14 mRNA levels were normalized to actin, and values relative to the 8-h time point for each strain are indicated in the black box. (C) Protein extracts from WT worms staged as in A were analyzed by Western blotting with antibodies against LIN-14 and control TUBULIN and ACTIN proteins. The LIN-14 protein levels were normalized to TUBULIN, and values relative to the 8-h time point are indicated in the black box. (D) The graph depicts the changes in lin-4 miRNA (triangles), lin-14 mRNA (diamonds), and LIN-14 protein (squares) over the 10-h time course in WT worms, using values calculated as described in A, B, and C.