Figure 1. 21UX-1 Targets the
xol-1 Transcript, Induces 22G-RNA Production, andRepresses
xol-1 Expression(A) 21ux-1 and 22G-RNAs targeting
xol-1 mRNA inwild-type, 21ux-1(deletion), 21ux-1(anti-gfp),
prg-1,
rde-3,
mago-12 mutants, and in HRDE-1/WAGO-9 input and IP samples are illustratedgraphically. Histograms represent the position
of50 end of 22G-RNAs at the
xol-1 locus. Theposition and base-pairing interaction between21ux-1 and
xol-1 are indicated above thegenome browser view. I marks A:U or G:C basepairs. + marks G:U wobble base pairs.(B) qRT-PCR of
xol-1 mRNA from total RNAs isolated from synchronized populations at late L4larvae and adult stages. Expression of actin mRNA(
act-3) served as the internal control. Data werecollected from three independent biological replicates. Error bars represent SD.(C) Representative differential interferencecontrast (DIC) image and fluorescence micrographs showing GFP::FLAG::XOL-1 expression(green) from wild-type and 21ux-1(deletion)animals. The dashed lines outline the position ofthe germline. Bright signals detected outside ofthe germline are from autofluorescence of gutgranules.(D) Western blotting analysis of proteins isolatedfrom gfp::flag::
xol-1 and 21ux-1(deletion)gfp::flag::
xol-1 strains at late L4 larvae and adultstages. The anti-FLAG antibody (top) detectingGFP::FLAG::XOL-1 and anti-tubulin antibody(bottom) were used. An asterisk marks a nonspecific band