Extended Data Fig. 6: Predicted UAD-2 IDRs are important for PRDE-1 foci formation and piRNA expression.
From: piRNA gene density and SUMOylation organize piRNA transcriptional condensate formation
a, IUPred2 prediction of UAD-2 and the USTC components using the IUPred2 long disorder option. b, qRT-PCR analysis of UAD-2 mRNA in wild-type animals and uad-2 deletion mutants. The levels were normalized to eft-3. Quantification of qRT-PCR data from n = 3 independent animals, presented as Mean ± SD. Statistical significance was determined using a paired-sample t test. Source data are provided as a Source Data file. c. Images showing the subcellular localization of the indicated UAD-2::GFP variants and mCherry::PRDE-1 in pachytene cells. All images were taken by the Leica THUNDER imaging System and deconvoluted using Leica Application Suite X software (version 3.7.4.23463). All images are representative of more than three animals (see also in Extended Data Fig. 6f). d, Graphs presenting the relative fluorescence unit intensity (RFU) of mCherry::PRDE-1 for wild-type animals and the indicated UAD-2 variants. The fluorescence intensity threshold is set to 8–255. Three worms were analyzed for each strain. Statistical significance was determined using a two-sample t test. Source data are provided as a Source Data file. e. Graphs presenting the relative fluorescence unit intensity (RFU) of wild-type animals and the indicated UAD-2 variants, marked by the dashed line. f. Images showing the subcellular localization of the indicated UAD-2::GFP variants and TOFU-4:mCherry in pachytene cells. g-j. Scatter plots displaying the second biological replicate expression levels of Cluster I, Cluster II, and out-Cluster piRNAs in the wild-type (x-axis) and indicated mutants (y-axis), based on the log2(reads per million+1). k. Boxplots revealing log2(piRNA reads per million+1) of the indicated adult animals of the second biological replicate. The box itself represented the interquartile range between 5% and 95%, thereby encompassing the middle 90% of the data set. The central horizontal line and the adjacent numeric value within the box represent the mean value. Error bars represent the mean ± 1.5 standard deviations (SD), indicating the variability of the data around the mean. Outliers are individually marked as points, representing data points that fall outside the range of mean±1.5 SD. Statistical significance was determined using a paired-sample t test.
