Fig. 2: GLH/VASA helicases are required for piRNA-dependent silencing of a piRNA reporter.

a GFP expression of the piRNA reporter in the indicated strains. Arrows indicate the adult germline nuclei in maturing oocytes with expression of GFP transgenes (top). DIC images of the corresponding reporter images (bottom). Bar, 10 micrometers. b Fluorescent micrographs show the localization of PRG-1 and WAGO-4 in the indicated strains in the adult germline (left) and in the four-cell embryo (right). Bar, 10 micrometers. c q-RT PCR measurements of the abundance of the GFP-targeting piRNA in the indicated strains. Statistical analysis was performed using a two-tailed Student’s t test. Bars indicate the mean, errors bars indicate the standard deviation, and data points indicate values for the three technical replicates from each genotype. d 22G-RNAs distribution at GFP coding sequences in the indicated strains. The green bar (below) indicates the location of the GFP sequence complementary to the GFP-targeting piRNA. e Photomicrographs of pachytene nuclei in fixed adult gonads hybridized with single molecule fluorescent (smFISH) probes complementary to gfp mRNA (Green) and nos-3 mRNA (Red) in the indicated strains. Nuclear DNA was stained with DAPI (Blue). The arrowheads indicate perinuclear mRNA foci while arrows indicate cytoplasmic mRNA foci. Bar, 5 micrometers. Photomicrographs are representative of data collected across three independent experiments. At least 10 gonads from each experiment were observed at random.