Fig. 5: EXD2-deficient cells engage BLM-dependent conservative mitotic telomere synthesis and form T-SCEs in a SMARCAL1- and MUS81-dependent manner.
a Immuno-FISH staining in WT and EXD2−/− U2OS synchronised prometaphase cells treated with siRNA targeting BLM, SLX4, SMARCAL1 or control siRNA. EdU incorporation was determined by Click-iT staining (red) and telomeres marked by a telomere-specific PNA FISH probe (TelG, green), DAPI (blue) acts as a nuclear stain (n = 80, 80, 76, 75, 55, 55, 51, 42, 54, 56, 62 and 48 prometaphases examined over 2 independent experiments, statistical significance was determined by two-sided Mann–Whitney analysis, bars represent +/− SEM, scale bar = 10 μm). b Quantification of conservative replication in U2OS WT and EXD2−/− as determined by two-replication round segregated CO-FISH using SpO and FITC-labelled PNA FISH probes specific to the telomeric repeat sequences. Cells treated with siRNA targeting SMARCAL1 or MUS81, or were treated with control siRNA targeting Luciferase, as indicated (n = 75 metaphases examined over 3 independent experiments, statistical significance was determined by two-sided Mann–Whitney analysis, bars represent +/− SEM). c Quantification of T-SCEs in U2OS WT and EXD2−/− as determined by one-replication round CO-FISH using SpO and FITC-labelled PNA FISH probes specific to the telomeric repeat sequences. Cells were treated with siRNA targeting SMARCAL1 or MUS81, or were treated with control siRNA targeting Luciferase, as indicated (n = 75 metaphases examined over 3 independent experiments, statistical significance was determined by two-sided Mann–Whitney analysis, bars represent +/− SEM). Source data are provided as a source data file.
