Fig. 4: NSUN2 depletion leads to stabilization of antisense DARTs.
From: NSUN2 facilitates DICER cleavage of DNA damage-associated R-loops to promote repair
a Heatmaps showing strand-specific chromatin-associated RNA sequencing (Chr-RNA-seq) read density in U2OS DIvA cells treated with (Z)−4-hydroxytamoxifen (+4OHT) to induce AsiSI-dependent DSBs. Left: sense-strand coverage in siCtrl and siNSUN2 conditions; right: corresponding antisense-strand coverage. Data are shown across ±2.5 kb flanking 80 well-characterized AsiSI cut sites (BLESS80). Each row represents one DSB site, ordered by increasing cleavage efficiency. Columns indicate genomic position relative to the DSB centre (0). Separate colour scales represent normalized signal intensity for each strand. b Box plot showing log2 fold change (siNSUN2 / siCtrl) in Chr-RNA-seq coverage for sense and antisense strands within ±500 bp flanking BLESS80 AsiSI-induced DSBs. c Metagene plots of Chr-RNA-seq coverage across ±2.5 kb flanking the BLESS80 DSBs. Top: sense-strand signal; bottom: antisense-strand signal, plotted as negative values for symmetry. Read coverage is shown for siCtrl (blue) and siNSUN2 (red) conditions. Values represent average per-nucleotide coverage across all sites for each strand. d, e Genome browser snapshots of strand-specific Chr-RNA-seq coverage at selected AsiSI sites located near the CREBZF (d) and PCGF1 (e) loci in U2OS DIvA cells. Data are shown for siCtrl and siNSUN2 conditions under +4OHT treatment. Strand-specific coverage is displayed for sense (red) and antisense (blue) transcripts. f, g Metagene plots of Chr-RNA-seq signal for sense (top) and antisense (bottom) strands across ±2.5 kb centred on two functional subsets of AsiSI-induced DSBs: (f) highly transcribed sites and (g) homologous recombination (HR)-prone sites. Strand-specific coverage is shown for siCtrl (blue) and siNSUN2 (red) cells after +4OHT treatment. Antisense signal is plotted as negative values to facilitate strand-resolved comparison. All data represent normalized, averaged RNA coverage per nucleotide across DSB subsets. Source data are provided as Source Data file.
