Fig. 4: TOPBP1 recruitment in response to rDNA breaks.
From: Treacle controls the nucleolar response to rDNA breaks via TOPBP1 recruitment and ATR activation
a Timecourse of TOPBP1 localization in GFP-Treacle expressing U2OS cells after I-Ppo1 transfection. b TOPBP1 localization 2 h after I-Ppo1 transfection in NBS1ΔN cells and NBS1ΔN cells complemented with wild-type and mutant NBS1-mNG (percentage of cells with > 2 TOPBP1 caps are indicated; one of two experiments is shown). c TOPBP1 localization 2 h after I-Ppo1 transfection in ATMi- and ATRi-treated cells versus vehicle (DMSO)-treated cells (percentage of cells with TOPBP1 foci inside of the nucleoli are indicated; one of two experiments is shown). d Quantification of cells with TOPBP1 caps after ATMi and ATRi treatment versus vehicle (DMSO) treatment (red bars represent mean). e Quantification of nucleolar EU incorporation after I-Ppo1 H98A (n = 692) and I-Ppo1 WT (n = 649) transfection in control U2OS and after I-Ppo1 H98A (n = 450) and I-Ppo1 WT (n = 521) transfection in siTOPBP1-treated cells (boxes represent the median with 25–75 percentile range and whiskers represent the 5–95 percentile range. Data points outside of this range are shown individually). f Endogenous Treacle localization 2 h after I-Ppo1 transfection in control siRNA and TOPBP1 siRNA transfected cells (percentage of cells with > 2 Treacle caps are indicated; one of two experiments is shown). g TOPBP1 localization in Treacle-depleted U2OS cells and control cells 2 h after I-Ppo1 transfection (percentage of cells with > 2 TOPBP1 caps are indicated; one of two experiments is shown). h HA-immunoprecipitations from 293FT cells transfected with the indicated HA-tagged Treacle variants, in the presence or absence of ATMi and ATRi and with and without IR treatment as indicated. STTT mutated Treacle: S171A, T173A, T203A, T210A. All scalebars = 10 µm. Source data are provided as a Source Data file.
