Extended Data Fig. 8: Local transcription enables NER irrespective of Mfd.

Supplementary to Fig. 5. a–h, Depriving the genomic loci of transcription drastically diminished NER irrespective of Mfd. Genomic DNA repair within the insulator was monitored as described in Methods for the lesions generated by 4-NQO (a, e), NFZ (b, f), cisplatin (c, g), or UV-C (d, h). lacZ was induced with IPTG followed by the exposure to drugs or UV radiation. Cells were allowed to recover for the indicated time intervals followed by the isolation of genomic DNA. Lesion density was determined by SLR-qPCR and used to calculate the percentage of repaired lesions. Values are the mean ± SD from 3 independent experiments. i, j, UvrAB recruitment to the UV-damaged DNA strictly depends on local transcription, but not Mfd (compare to Fig. 5l, m). Recruitment of UvrA (i) and UvrB (j) to the lacZ insulator (with or without promoter) was determined by ChIP–qPCR in Δmfd cells as in Fig. 5h. Results are shown as a fold change in the occupancy of UvrAB within the insulator of Δmfd following UV irradiation. **P < 0.01, ****P < 0.0001. Statistical analysis was performed using unpaired non-parametric two-tailed Mann-Whitney t test. Values are the mean  ± SD from 3 independent experiments.