Fig. 2: USP50 promotes replication in unperturbed and stressed conditions.

Where included, graphs indicate the mean ± SEM, exact P values are shown, and number of biological repeats is listed (n). Other than 2H which was two-way ANOVA, all statistical analysis in this figure was performed using a two-tailed unpaired t test. Source data are provided with this paper. A Mean% of first-label terminations after non-targeting control (NTC) siRNA (−) or shUSP50 (+) and complemented with FLAG-USP50 (left), I141R-FLAG-USP50 (right) or uninduced (−). n = 3, >195 fibres per condition. B Mean ratio of second-label tracts either side of first label after treatment as in A. n = 3, >35 first-label origins per condition. C Mean% of stalled forks after treatment as in A. n = 3, >240 fibres per condition. D Immunoblot of RPA, pRPA and vinculin following 3 hours 5 mM HU. Quantification shows mean pRPA from n = 5. E Native BrdU tracts length after treatment as in A and with HU. n = 3, >1400 tracks per condition. F 53BP1 foci numbers after treatment as in A. n = 3, >150 cells per condition. G 53BP1 foci numbers after treatment as in A. n = 2, >100 cells per condition. H Colony survival after treatment as in A and 16 hours HU. n = 4. I Colony survival after siNTC or shUSP50 and 24 hours Pyridostatin. n = 2. J 53BP1 foci numbers after siNTC (−) or shUSP50 (+), with or without 24 hours 100 µM Pyridostatin. n = 3, 250 cells per condition. K GC content of break-adjacent heximeric sequences enriched or reduced in USP50:siNTC treated cells from overlapping sequences between n = 2 biological repeats. Occ-p: p value of the occurrence difference. Statistical test: hypergeometric for over/under-representation. All significant sequences and p values shown in Supplementary Fig. 4B.