Fig. 3: Widespread reduction in canonical miRNA levels in pash-1(ram33[G179R]) mutants.

a, b Log₂ average geometric-mean normalized sRNA-seq read counts in wild-type (x-axis) and pash-1(ram33[G179R]) (y-axis) animals grown at 20 °C (a) or 25 °C (b). Small RNA features are represented by data points colored by their classification. Diagonal lines show 0-, 2-, and -2-fold enrichments. The Wald test was used for statistical analysis. The inset bar plots show the numbers of miRNAs represented by >20 geometric mean-normalized reads significantly down or upregulated (p < 0.05) or unchanged (p ≥ 0.05) in pash-1(ram33[G179R]) relative to wild-type (no fold-change cutoff was applied). n = 3 (a) or 4 (b) biological replicates per strain. c Classification of miRNAs upregulated (p < 0.05) in pash-1(ram33[G179R]) relative to wild-type animals grown at either 20 °C or 25 °C, as indicated, based on data in (a) and (b). d Secondary structure predictions of miR-1829 family miRNA hairpins. Nucleotide sequences are shown for the miRNA duplex regions. The cleavage sites are indicated with scissors. e Mean reads per million (rpm)-normalized sRNA-seq counts for iso-miRs (offset by ±1–3 nt relative to miRNA 5’ end) and small RNAs derived from pre-miRNAs (pre-miRNA derived reads offset at their 5’ ends by >3 nt relative to mature miRNA 5’ end) in wild-type and pash-1(ram33[G179R]) mutants grown at 20 °C or 25 °C. Data as in (a) and (b). Error bars are SD. n = 3 (20 °C) or 4 (25 °C) biological replicates per strain. Two-tailed, two-sample Student’s t-tests were used to calculate p-values for comparisons to wild-type. A Bonferroni correction for two comparisons was applied to each. Source data are provided as a Source Data file.