Figure 2. Transgenerational Inheritance Requires NRDE-2 and the Germline-Specific Nuclear Argonaute HRDE-1/ WAGO-9 (A) Biosorter analysis of WT,
nrde-2, and
hrde-1/
wago-9 animals showing the failure of heritable silencing in these mutant strains. GFP fluorescence of the transgene and the percentage of silenced animals per plate were determined using a large particle biosorter and FlowJo. Ten silenced worms were selected where possible from each plate to produce the next generation. At least 500 worms were analyzed per plate with the following number of replicates (empty vector, GFP RNAi, F1, F2, respectively). WT, n = 7, 6, 4, 4;
nrde-2, n =3, 3, 3, 3;
hrde-1/wago-9, n = 12, 6, 6, 6. Error bars represent the SEM. (B) HRDE-/WAGO-9 is expressed in the germline. Wild-type dissected germlines (adults) were stained with anti-HRDE-1/WAGO-9 (green) and a P-granule-specific antibody (OIC1D4, red). DNA was costained with DAPI (blue). Images on the right are merged from all three channels. (C and D) HRDE-/WAGO-9 is a nuclear protein. Immunostainings were performed on dissected gonads from adult wild-type (N2) or
hrde-1/wago- 9 (
tm1200) animals using anti-HRDE-1/WAGO-9 (green) and anti-a-tubulin antibodies (red). DNA was costained with DAPI (blue). Images on the right are merged from all three channels. Images shown are germ cells in the transition zone/ pachytene region (C) and oocytes (D).