Fig. 7: C16orf72 and BRCA1/Senataxin function together to promote replication fork restart and tolerance to replication stress.

a Parental and c16orf72Δ U2OS cells were transfected with control (siCTRL), BRCA1, or Senataxin (SETX) siRNA. Following treatment of cells with CIdU, HU and IdU as illustrated (top panel), DNA fibres were prepared from cells and quantified for the CIdU/IdU staining. 300-900 DNA fibres were analysed per condition from n = 3. Fork restart represents the % of fibres that display both CldU and IdU staining (upper panel). Data are presented as mean values +/− SEM.  b, c Clonogenic survival assay of parental and c16orf72Δ U2OS cells transfected with control (siCTRL), Senataxin (siSETX; b) or BRCA1 (siBRCA1; c) siRNA following exposure to increasing concentrations of HU. Data represent 2 biological repeats. Data are presented as mean values +/− SEM. d Parental and c16orf72Δ U2OS cells treated with DMSO or Olaparib as indicated prior to immuno-fluorescence with the S9.6 antibody. Data represents the quantification of nuclear mean intensity of the S9.6 signal. Mean values (red lines) are represented +/− SEM where at least 173 cells were examined per treatment over 3 biological independent experiments. e, f Clonogenic survival assay of parental (WT) and c16orf72Δ U2OS cells transfected with control (siCTRL), BRCA1 (siBRCA1; e) or Senataxin (siSETX; f) siRNA following exposure to increasing concentrations of olaparib. Data are presented as mean values +/− SEM where n = 5 independent biological repeats for (e) and n = 3 independent biological repeats for (f). In the case of DNA fibre and S9.6 staining, statistical significance was assessed by Ordinary one-way ANOVA or Kruskal Wallis non-parametric test. Clonogenic survival assays were assessed by two-way ANOVA (*p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001 and ns = not significant). Source data are provided as a Source Data file.