Fig. 6: Ribosome occupancy affects CSR-1-mediated maternal mRNA clearance.

a Schematic of the germline-expressed single-copy mCherry::h2b transgenic mRNA fused to a 3′UTR derived from an early- (egg-6 3′UTR) or a late-degraded (tbb-2 3′UTR) mRNAs. b RT-qPCR assay to detect the levels of mCherry mRNA from the two transgenic reporters in adults and early embryos. The bars indicate the mean, the error bars the standard deviation, and the black dots individual data from three or two biologically independent replicates in adults and embryos respectively. The levels of mCherry mRNA are normalized on the levels of ribosomal 18S RNA. c Translational efficiency (TE) of mCherry::h2b transgenic mRNAs. The lines indicate the mean and the dots individual data from two biologically independent experiments. d Abundance of CSR-1-bound 22G-RNAs antisense to mCherry (left) or the coding sequence of embryonic CSR-1 targets (right). The lines indicate the mean and the dots individual data from two biologically independent experiments (left). For the box plots (right), the line indicates the median value, the box indicates the first and third quartiles, and the whiskers indicate the 5th and 95th percentiles, excluding outliers. Two-tailed P value were calculated using the Mann–Whitney–Wilcoxon test. Sample size n = 2351 (CSR-1 mRNA targets). e Metaprofile analysis showing normalized 22G-RNA reads (RPM) across mCherry nucleotide sequence from mCherry::h2b transgenic mRNA fused to egg-6 3′UTR (left) or tbb-2 3′UTR (right) in CSR-1 immunoprecipitation (IP) or total RNA input. The average from two biologically independent replicates is shown. Source data are available online.