TRUB1-mediated pseudouridylation occurs in the nucleus. (A) HEK293T cells overexpressing FLAG-tagged TRUB1 were stained with αTRUB1 (red), αFLAG (green), and DAPI (blue). Representative image (600×) is shown, with overexpressed TRUB1 found in both the cytoplasm and the nucleus of transfected cells. (B) Depiction of aggregate Ψ-ratios across the SGSM2 gene in the cytoplasmic-enriched (top) or nuclear (bottom) fractions; values at pseudouridylated position 2412 are highlighted in yellow. (C) Scatterplot depicting aggregate Ψ-ratios across the nuclear and cytoplasmic fractions for 20 sites that passed the thresholds of detection in our pipeline, harbored a “GUUC” core motif, and with ≥15 reads overlapping the site in the treated fraction. The regression line (lm[y ∼ x]) and the y = x lines are depicted.
