Fig. 5: PGL-1 assembly is required for tethered mRNA reporter repression.
From: C. elegans germ granules require both assembly and localized regulators for mRNA repression
a Protein–mRNA tethering assay and PGL-1 assembly. To test the necessity of granule formation for mRNA repression, NtDD assembly mutations were added to PGL-1::SNAP::λN22 and germlines observed for GFP reporter expression. N-terminal dimerization domain (NtDD, yellow), central dimerization domain (CDD, orange), SNAP (magenta), λN22 (light blue) and RGG repeats (blue), and GFP (green). Modified from76. b–e GFP reporter expression in germ cells of live animals. (b, c) Brightfield image. (d, e) GFP fluorescence (green); auto fluorescence (red). n, number of animals scored for GFP expression. Scale bar, 10 μm, in (b) applies to (b–e) images. f–i Representative images of PGL-1 granule formation, seen by SNAP staining (magenta) and GFP fluorescence (green) in fixed gonads. DNA (DAPI) in cyan. n, number of germlines scored for GFP expression. %, germlines with detectable GFP. Fisher’s exact test of PGL-1::SNAP::λN22 vs. PGL-1 (K126E K129E)::SNAP::λN22 (p-value < 0.0001). Scale bar, 10 μm, in (f) applies to (f–i) images. Figure 1 and Fig. 5 results were performed in parallel, and thus results from (b, d, f, h) are the same as reported in Fig. 1e, g, i, k.
