Fig. 5: Replication defects in USP50 deficient cells are driven by RECQL4/5.

Where included, graphs indicate the mean ± SEM, exact P values are shown, and number of biological repeats is listed (n). 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 stalled forks after shUSP50 with RECQL4 siRNA. n = 3, >200 fibres per condition. B Mean% of stalled forks after shUSP50 with RECQL5 siRNA. siNTC and shUSP50 conditions are shared with 5 A. n = 3, >200 fibres per condition. C Mean% of first-label terminations after shUSP50, with or without RECQL4 or RECQL5 siRNA. n = 3, >200 fibres per condition. D Native BrdU tracts length after shUSP50 and siRNA targeting RECQL4 and RECQL5 or DNA2 and 3 hours 5 mM HU. siNTC and shUSP50 conditions are shared with 2E. n = 3, >1400 tracks per condition. E Mean 53BP1 foci after shUSP50 and RECQL4 and RECQL5 siRNA and 3 hours 5 mM HU followed by 20 mins recovery. n = 3, 150 cells per condition. F Mean% of stalled forks after FEN1 siRNA with and without siRNA targeting RECQL4 and RECQL5. n = 3, >400 fibres per condition. G Mean RECQL4 foci after shUSP50 and 3 hours 5 mM HU, followed by 20 mins recovery. Representative images left, scale bar is 10 µm. n = 3, 150 cells per condition. H Mean PLA foci of RECQL4 with Biotin-EdU after RECQL4 siRNA, or shUSP50 in cells pulsed with EdU for 15 mins and 3 hours 5 mM HU, or HU treated, washed and allowed to recover for 20 mins (HU release). n = 3, >150 cells per condition. I Mean PLA foci of RECQL4 with Biotin-EdU after RECQL5 siRNA, or shUSP50 in cells pulsed with EdU for 15 mins and 3 hours 5 mM HU, or HU treated, washed and allowed to recover for 20 mins (HU release). n = 4, >200 cells per condition. J Colony survival after shUSP50 and siRNAs to DNA2 or RECQL4 and RECQL5 and 24 hours 25 μM Pyridostatin. n = 7 for siNTC and shUSP50, n = 3 for all other conditions.