Extended Data Fig. 9: Expression and TCR activity of UVSSA mutants.
a. Expression of the indicated GFP-UVSSA proteins in RPE1-iCas9 UVSSA-KO cells. The data shown represent at least three independent experiments. Scale bar, 15 μm. b. Western blot analysis of GFP-UVSSA proteins (or empty control) in RPE1 UVSSA-KO cells. The data shown represent at least three independent experiments. c, d. Measuring transcription recovery (RRS) by 5-ethynyl-uridine labelling in the indicated RPE1-iCas9 cells following UV irradiation (3 h or 24 h; 12 J/m2). Representative images (c) and quantification (d) of 5-EU levels normalized to mock treatment for each cell line. The experiment was performed three times. Each coloured circle represents 1 cell. Each black circle represents the mean of two technical replicates, with >80 cells collected per technical replicate. The black lines represent the mean of all three independent experiments. e, f. Measuring TCR-dependent γH2AX induction following trabectedin exposure in replicating cells labelled with 5-ethynyl-deoxyuridine (EdU) in the indicated RPE1-iCas9 cells. Representative images (e) and quantification (f) of γH2AX levels normalized to trabectedin-treated WT cells within each experiment. The experiment was performed three times. Each black circle represents the mean of two technical replicates, with >70 cells collected per technical replicate. The black lines represent the mean of all three independent experiments. Scale bar, 10 μm.
