Extended Data Fig. 1: Characterization of Slx4∆125/∆ cells.
a. RT-qPCR analysis of Slx4 mRNA normalized to Gapdh mRNA using the 2–ΔCT method in three independent experiments. Each data point is an average of three technical replicates. Data shows mean of three independent biological replicates (n = 3), Error bars: standard deviation (s.d.) Statistical analysis using Student’s t-test. b and c. Ratio of Tus/Ter-induced (b) and I-SceI-induced (c) LTGC: total HR in Slx4+/+ clones and Slx4∆125/∆ clones. Data shows mean values, n = 6. Error bars: s.e.m. Analysis by ANOVA. P-value *p < 0.05 and ****p < 0.0001 d. CRISPR/Cas9 with dual sgRNA targeting to generate Slx4+/– hemizygous cells. The Slx4– allele contains a 19.3 kb deletion between exons 2 and 15. Red half arrowheads: PCR and sequencing primers specific to Exons 2 and 15. Predicted PCR product sizes for Slx4+ (wild type) allele shown. e. Gel shows PCR products detected using gDNA from Slx4+/+ and Slx4+/– clones. Note the 444 bp PCR product formed across the 19.3 kb deletion between exons 2 and 15 in Slx4+/– cells. f. DNA sequencing chromatogram of PCR products from (e). g. RT qPCR analysis of Slx4 mRNA in Slx4+/+ and Slx4+/– cells. Data shows mean of three independent experiments (n = 3) normalized to Gapdh mRNA using the 2–Δ∆CT method and analyzed by unpaired Student’s t-test. Error bars: s.d.. h. and i. Quantification of colony formation of Slx4+/+ vs. Slx4+/– clones in the presence of MMC (h), Olaparib (i). Data shows mean of three biologically independent replicates, n = 3. Error bars: s.d. j. Tus/Ter-induced and I-SceI-induced repair frequencies in Slx4+/+ and Slx4+/– cells. Data shows mean values of five biologically independent replicates, n = 5. Error bars: s.e.m. Statistical analysis using ANOVA.
