nrde-3-dependent silencing of endogenous RNAi targets and transgenes.

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nrde-3-dependent silencing of endogenous RNAi targets and transgenes. nrde-3-dependent silencing of endogenous RNAi targets and transgenes. (A) eri- and nrde-3-dependent changes in relative mRNA levels for select endo-siRNA target genes. Log2 ratio (relative to N2 = 0) of qPCR determined mRNA levels for the indicated genes from L4 larvae of the indicated genotypes. Expression of these five genes, which are targets of rrf-3-dependent endo-siRNAs, was previously reported to be up-regulated in rrf-3 mutants (Gent et al. 2010). Transcript abundances are normalized to gpd-3. P-values are calculated by t-test; *P < 0.05; **P < 0.01. (B–G) Representative photomicrographs of sur-5p::gfp expression in (B) wild type, (C) nrde-2(gg91), (D) nrde-3(gg66), (E) eri-1(mg366), (F) eri-1(mg366); nrde-2(gg91), and (G) eri-1(mg366); nrde-3(gg66) strains. The extensive transgene silencing that is readily apparent in intestinal cells (yellow arrows) in the eri-1(mg366) background is dependent on nrde-2 and nrde-3. Compared to nrde-2(gg91) and nrde-3(gg66), wild-type animals show reduced gfp levels. (H) Average gfp fluorescence of hatch-synchronized 24-hr adult (20 °C) worms. For each genotype the average whole body fluorescence (0.1-sec exposure) of 10 worms is shown. Error bars represent standard deviation. P-values are calculated by t-test; red asterisks are P < 0.01 compared to wild type (N2) and blue asterisks are P < 0.01 compared to eri-1(mg366). Jimmy J. Zhuang et al. Genetics 2013;194:117-131 Copyright © 2013 by the Genetics Society of America