Figure 1.
rde-12 Is Required for RNAi and Is Localized to P Granules(A) Gene structure of
rde-12. Exons are depicted in boxes. Mutant alleles and corresponding changes in the protein sequence are indicated.(B) Relative abundance of endogenous
unc-15 mRNA in synchronized wild-type (WT),
rde-1(
ne300),
rde-12(
hj41), andrde-12
(hj41); hjSi394[
rde-12(+)] animalssubjected to
unc-15 RNAi compared to no RNAi control (L4440 vector). The mean + SD from three independent samples assayed in triplicates are shown.(C)
rde-12(
qt131) animals were sensitive to RNAi against
pal-1 when high doses of dsRNA were directly injected into the gonad. The number of injected animals is n = 8-17. Mean 6 SD is shown. At 100 and 50 ng/ml, wild-type and
rde-12 mutant animals were not significantly different. At 10 ng/ml, p < 0.01.(D-G) The germline (in the loop region connecting the proximal and distal gonad arms) of live animals was imaged by confocal microscopy. The inner borderof gonad arms is depicted by dotted lines. Full-length RDE-12 was fused to GFP (GFP::RDE-12). (D) mRuby::PGL-1, (E) tagRFP::RSD-6, (F) mRuby::DCAP-1,and (G) MUT-14::mCherry are shown. Solid arrowheads indicate colocalization of GFP with RFP signals. Open arrowheads indicate the positions of GFPsignals that do not overlap with RFP signals. Boxed areas magnified 33 are shown in the far-right panels. Scale bars, 6.5 um.