Figure 3: RSD-3 is required for importing silencing RNA into cells. | Scientific Reports

Figure 3: RSD-3 is required for importing silencing RNA into cells.

From: Endomembrane-associated RSD-3 is important for RNAi induced by extracellular silencing RNA in both somatic and germ cells of Caenorhabditis elegans

Figure 3

(a) rsd-3(tm9006);ccIs4251 (myo-3p::gfp) animals show resistance to GFP feeding RNAi. Body wall muscle GFP fluorescence in ccIs4251 animals is efficiently reduced by GFP RNAi food (lower left panel), but not in rsd-3(tm9006);ccIs4251 (lower right panel). Scale bars, 50 μm. (b) Body wall muscle-specific expression of RSD-3 restores GFP feeding RNAi sensitivity in body wall muscles of rsd-3(tm9006);ccIs4251. rsd-3(tm9006);ccIs4251;Ex[myo-3p::rsd-3 & myo-3p::mCherry] worms were fed with mock or GFP RNAi food. myo-3p::mCherry indicates the cells expressing RSD-3. Arrows indicate the cells in which RSD-3 is expressed and GFP RNAi is restored. Arrowhead indicates the cell in which RSD-3 is not expressed and GFP RNAi defect is not rescued. Scale bars, 20 μm. (c) Intestine-specific expression of RSD-3 does not restore GFP feeding RNAi sensitivity in body wall muscles of rsd-3(tm9006);ccIs4251. rsd-3(tm9006);ccIs4251;Ex[ges-1p::rsd-3 & ges-1p::DsRed] worms were fed with mock or GFP RNAi food. DsRed fluorescence confirms the expression of rsd-3 in the intestine (lower panels). Scale bars, 20 μm. (d) rsd-3(tm9006) animals show partial resistance to pseudocoelomic injection of pos-1 dsRNA. pos-1 dsRNA (10 ng/μl) were injected into the pseudocoelom of more than twenty wild type and rsd-3(tm9006) animals, and percentage of hatched progeny was scored for each injected animal. Data is shown as mean ± SEM. *P < 0.05 (Student’s t-test, two-tailed).

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