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. 2010 Sep 2;6(9):e1001089. doi: 10.1371/journal.pgen.1001089

Figure 5. Northern blot results for mir-57 overexpression.

Figure 5

(A) 20 µg total RNA from wild type (wt) or mir-57 mutant (Δ) L4 worms with (+) or without (-) injection of genomic DNA containing both regulatory and structural sequences of mir-57 or with the injection of mir-57 promoter only were separated on a 15% urea denaturing gel, blotted and hybridized using an antisense mir-57 LNA probe and an antisense U6 DNA probe (See Materials and Methods). In order the lanes show wild type; the mir-57 deletion strain; animals with transgenic arrays of the mir-57 genomic fragment in the wild type background; animals with transgenic arrays of the with mir-57 genomic fragment in the mir-57 deletion background; and finally animals with arrays of the mir-57 promoter in a wild type background. Arrays of the mir-57 genomic fragment into both wild type and the mir-57 deletion mutant resulted in elevated levels of the mir-57 miRNA. Arrays of mir-57 promoter alone in the wild type background produced mir-57 miRNA levels similar to that seen using the intact mir-57 region. No signal was detected for mir-57 deletion strain. (B) Quantification of expression changes measured by the intensities of the mir-57 signal from lanes in panel A and normalized against U6 (See Materials and Methods).